Bio-insecticidal potential of amylase inhibitors

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458

Review Article ISSN: 2321-4988

Available online through http://jprsolutions.info

Bio-insecticidal potential of amylase inhibitors Parag Gupta1, Anuradha Singh1, Gaurav Shukla2, Neeraj Wadhwa* 1, 2

Department of Biotechnology, Jaypee Institute of Information Technology, Noida,Uttar Pradesh, India. Received on:16-01-2013; Revised on:19-02-2013; Accepted on:24-03-2013

ABSTRACT Agricultural crops are facing several losses due to pest. Pesticides, biopestcides , plant defence compounds are different ways to protect the agricultural crop losses. The enzyme inhibitors act on insect gut digestive α-amylases and proteinases, which play a key role in the digestion of plant starch and proteins. Starchy leguminous seeds are an important staple food in manycountries. These seeds are rich in protein, and carbohydrate therefore suffer extensive predation by bruchids (weevils)and other pests. Insects like weevils are highly dependent on starch for their energy supply. In this review, we summarize the effects of pesticides, need of bio pesticides, different types and sources of plant αamylase inhibitors their interaction with insect α-amylases and subsequent insect mortality. Keywords: α-amylase inhibitors, knottin-like; lectin-like; thaumatin-like; Kunitz; cereal-type; Pesticides, Bio pesticides, pest, TMA INTRODUCTION Pests are responsible for several agricultural damages. It is estimated that 37% of rice, 26% of soybean, 40% of potatoes, 31% of maize and 28% of wheat are lost because of pest attack and infestation [1, 2]. Maximum damage is caused to starchy food products which are an important staple food to the pest. Several insect feed on starchy seeds during larval and/or adult stages; depend on endogenous α-amylases for their survival.

be targeted to control the insect growth and thereby protect the stored grain. Strategies of pest control:

1. Use of Pesticides: Agricultural pesticides are used in the cocoa, coffee and cotton farming, in vegetable and fruit production, and for other mixed crop farmα− Amylase is present in digestive systems of insects belonging to ing systems involving cereals (mostly maize), tuber crops (e.g. yam, order Orthoptera, Diptera, Hymenoptera, Lepidoptera and Coleoptera cassava), legumes (e.g. cowpeas, beans), sugarcane, rice, etc. [3]. Since the metabolic energy requirement is essential for larval development this energy is received by starch hydrolysates[4]. Insect Pesticides are unique, intrinsically toxic chemicals designed to be pest like Rhyzopertha dominica, cowpea weevil,(Callasobruchus deliberately spread into the environment to kill off pests. Insect-pest maculates) seed weevils and maxican bean weevil (Zabrotus control largely depends on application of organochlorine insecticides, subfaciatus), Red flour beetle, Tribolium castaneum (Herbst) are which have low efficiency and are extremely hazardous not only to extensively starch dependent insects and utilize α-amylase for their farmers and consumers, but also to the environment [10]. The costs survival [5-9]. α-Amylases (α-1,4-glucan-4-glucanohydrolases, of pesticides, estimated at more than US $10 billion per annum, and EC 3.2.1.1) constitute a family of endo-amylases that catalyses the they often affect non-target organisms and leave harmful residues hydrolysis of α-d-(1>4)-glucan linkages in starch components, gly- [11]. Commonly used insecticides in developing countries mainly cogen and other carbohydrates. The enzyme plays a key role in car- consist of organochlorines (DDT, endosulfan, lindane, dieldrin), orbohydrate metabolism of microorganisms, plants and animals. ganophosphates (monocrotophos, parathion, methamidophos) and carbamates (carbofuran, thiodicarb, maneb) noted for their toxicity These enzymes catalyze the initial hydrolysis of starch into shorter [12] oligosaccharides that could be assimilated by organisms. Due to prominent digestive and survival role of α-amylase in insects it can 1a. Drawbacks of Use of Pesticide: *Corresponding author. Dr. Neeraj Wadhwa Department of Biotechnology, Jaypee Institute of Information Technology, A-10, sec 62, Noida, Uttar Pradesh, India.

Long term application of pesticides results in adverse effects on the beneficial organisms, leaves pesticide residues in the food and results in environmental pollution and harmful effects on humans like cancer and neurodegenerative diseases [13-15], respiratory effects [16, reproductive and developmental toxicity [17] and altering sperm

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 nuclear proteins [18]. The presence of pesticide residues in fruits and maculates and Zabrotes subfasciatus , T. castaneum, R. dominica vegetables can be a significant route to human exposure and most of [33-38]. organochlorine pesticides have been banned because they are highly persistent insecticides, and their residues still appear as pollutants in α -Amylase inhibitors, depending upon the number of aminoacid food as well as in the environment. Some examples of persistant pes- residues and cysteine content have been classified into six different ticides reported from food used for human consumption are enlisted classes according to their tertiary structure: including lectin-like, in Table 1 knottin-like, cereal-type, Kunitz-like, γ-purothioninlike, and thaumatinlike [30]. The specificity of these inhibitors depends on the type and Table 1 Some persistant pesticides and along with their sources and source of enzyme. harmful effect Alpha amylase inhibitors are extensively found in many plant seeds Source Pesticide residue Harmful effects Reference and tubers and are abundant in cereals [39-42] and legumes [37, 4345]. Proteinaceous inhibitors are mainly present in cereals such as Wheat Mercaptothion C E N [19-22] wheat Triticum aestivum [37,45,46], barley Hordeum vulgareum [47], Permethrin C E Cypermethrin C E R N sorghum Sorghum bicolor [48], rye Secale cereal [49,50] and rice Chlorpyrifos C E R N Oryza sativa [44] but also in leguminosae such as pigeonpea Cajanus Fenitrothion E N cajan [41], cowpea Vigna unguiculata [42] and bean P. vulgaris [40,51]. Fenvelerate E R Proteinaceous α-amylase inhibitors have also been identified in root Proprioconazol C R DDT C crops and they have been purified and characterized from the tubers Cis-Chlordane of Colocasia and Dioscorea alata [52- 55]. The natural defenses of crop plants can be improved through the use of transgenic technolTomatos Malathion C E N [23] ogy. Pepper Omethioate Legumes

Chlorpyrifos

C

E

R

N

[22,24-26]

Pakchoi cabbage

Cypermethrin

C

E

R

N

[22,24-26]

Rice

chlorpyrifos

C

E

R

N

[18,22,25-29]

Apple

Methamidophos

E

R

N

(C= Carcinogenic, E= Endocrine disruptor, R==Reproductive and developmental toxin, N= Neurotoxin)

2. Plant defense compound: Plants have evolved a certain degree of resistance to herbivores and pathogens through production of defence compounds, which may be toxic secondary compounds(e.g. antibiotics, alkaloids, terpenes, cyanogenic glucosides) and protein antimetabolites(e.g. chitinases, ß-1,3-glucanases, lectins, arcelins, vicilins, systemins, enzyme inhibitors such as protease inhibitors and α-amylase inhibitors [30]. Host plant resistance and natural plant products offer a potentially better economical and safe method for insect pest control. They are safe to the non-target beneficial organisms and human beings [31].Some plants don’t have their own effective resistance mechanism like wild varieties of wheat and barley therefore plant traits that contribute to pest resistance need to be reinforced using new approaches. New control methods to diminish reliance on insecticides are α-amylase inhibitors, protease inhibitors, lectins and possibly δendototoxin [32]. Plant α-amylase inhibitors have been reported in growth, development, and reproduction of insect pest.The effect of proteinaceous inhibitors on insect development was observed by in vitro insect trials on C. cephalonica, Acanthoscelides obtectus, Callosobruchus

3. BIOPESTICIDES; NEED OF THE HOUR Biopesticides or biological pesticides are relatively low-risk pesticides derived from such natural products that do not cause harm to human beings and control target pests by a specific mechanism. Biopesticides include naturally occurring chemicals and living organisms used in insect, weed and disease control. Biopesticides based on living microbes and their bioactive compounds have been researched and promoted as replacements for synthetic pesticides for many years. Advantages of biopesticides over chemical pesticides: • Biopesticides are more specific • Fewer negative environmental impacts, makes them suitable for use in urban or sensitive environments. Limitations of biopesticides • Cost issues • Lack of efficacy • Inconsistent field performance • Limited availability of biopesticides. • Limited persistence and efficacy in comparison with chemical pesticides. [56] Recently, technological advances and major changes in the external environment have positively altered the outlook for biopesticides. In spite of the success of the introduction into a tobacco plant of the genetic material coding for a entomotoxic protein from the bacterium Bacillus thuringiensis. There are difficulties for acceptance of these ‘anti-natural’ products by the consumers and some concerns about its bio-safety in mammals. [57- 60] Significant increases in market penetration have been made, but

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 4. CHOICE OF α -AMYLASES INHIBITOR An alternative strategy could be to take advantage of the plant’s own defense mechanisms , by manipulating the Table 2 Plant inhibitors sources with targeted insect (literature review till 2012) expression of their endogenous defense proInhibitor Name Source Insect Targeted Reference teins like are lectins, arcelins, chitinases, b1,3glucanases, defensins and digestive enzyme SIa1, SIa2 S. bicolor Locusta migratoria [48] inhibitors or introducing an insect control and SIa3 Periplaneta americana A-1 Colocasia esculenta Bacillus species [53] gene derived from another plant.[61-65]

biopesticides still only make up a small percentage of pest control products.

B-2 α AI-Pc1 PAI Colocassia a-amylsae Inhibitor

Colocasia esculenta Phaseolus coccineus C. cajan corms of Colocasia

CpAI Wheat α- amylase Inhibitor T-αAI AAI

Carica papaya seeds Wheat

Secale cereale α- amylase Inhibitor αAI-Pa1

Secale cereal(rye)

BIII Cerrado Plants Inhibitors

Triticale seeds A. hypochondriacus

Phaseolus acutifolius A. Gray S. cereale Cerrado plants

α-AI1

Annona crassiflora leaf Aspidosperma macrocarpon Calophyllum brasiliense Kielmeyera coriacea Qualea grandiflora Qualea parviflora P. vulgaris

α-AI2 0.19

P. vulgaris T. aestivum

0.53

T. aestivum

0.28 Wheat Extract Inhibitor

T. aestivum T. aestivum

1,2 and 3 CAI WRP25

S. cereale V.unguiculata T. aestivum

WRP26

T. aestivum

WRP27

T. aestivum

Zeamatin

Z mays

Bacillus species Hypothenemus hampei Helicoverpa armigera (low) Callosobruchus chinensis Tribolium castaneum Corcyra cephalonica Spodoptera littoralis Callosobruchus maculates Rhyzopertha dominica Eurygaster integriceps Tenebrio molitor Hypothenemus hampei Prostephanus truncates Acanthoscelides obtectus Zabrotes subfasciatus Callosobruchus maculatus, Zabrotes subfasciatus Zabrotes subfasciatus Acanthoscelides obtectus Zabrotes subfasciatus Acanthoscelides obtectus

Callosobruchus maculates Callosobruchus chinensis Hypothenemus hampei Tenebrio molitor Diabrotica virgifera Zabrotes subfasciatus Acanthoscelides obtectus Diabrotica virgifera Zabrotes subfasciatus Sitophilus oryzae Tenebrio molitor Callosobruchus maculates Zabrotes subfasciatus Tenebrio molitor Callosobruchus maculatus Acanthoscelides obtectus Tenebrio molitor Lygus lineoralis Lygus Hesperus Diabrotica virgifera Tenebrio molitor Callosobruchus maculates Sitophilus oryzae Tribolium castaneum Callosobruchus maculatus Zabrotes subfasciatus Tenebrio molitor Sitophilus oryzae Tribolium castaneum Callosobruchus maculates Tenebrio molitor Sitophilus oryzae Tribolium castaneum Rhyzoperta dominica Sitophilus zeamais

[77] [41] [35]

[78] [38] [79] [68,80,81] [49] [49,82,83]

[84]

[68, 85,86]

[9,87] [37,45,88]

[45,88]

[88] [37] [50] [42] [37,45]

[ 37,45]

[37] [89]

BioMedRx Vol.1 Issue 5 .May 2013

α-Amylase inhibitors occur in many plants as part of the natural defense mechanisms. amylase inhibitors are of great interest as potentially important tools of natural and engineered resistance against pests in transgenic plants [66,67,68] Particular attention has been focused on the lectin-like inhibitors present in the common bean P. vulgaris seeds, which have been shown to have toxic effects to several insect pests. The effect of the bean amylase inhibitors on the amylases of different organisms was well determined not only by enzymatic activity, but also in feeding assay experiments [40, 69-71]. Complete resistance against bruchids, the pea weevil (Bruchus pisorum ), the cowpea weevil (C. maculatus ) and the azuki bean weevil (Callosobruchus chinensis ), was found in transgenic pea and azuki bean seeds expressing the inhibitor, aAI-1, of the domesticated common bean P. vulgaris [7174]. The transgenic grains showed minimal effects on mammalian digestion system [75] suggesting that these proteins can be safely introduced into food plants. 4A. Mode of action of enzyme inhibitors and their specificity Most of the inhibitors are proteins that inhibit the enzymes by forming a complex by blocking the active site of enzymes which finally leads to reduction in its catalytic activity [76]. Different α-amylase inhibitors from cereals and beans (Phaseolus vulgaris) have different molecular structures, leading to different modes of inhibition and different specificity profiles against a diverse range of aamylases. Specificity of inhibition is an important issue as the introduced inhibitor must not adversely affect the plant’s own α-amylases or human amylases and the nutritional value of the crop. 4B. Mechanism of action: a - amylase inhibitors have potential applica449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 tion in various fields namely treatment of diabetes to crop protection. Some general aspects of the inhibition strategy are as follows; The inhibition occurs mainly via interactions within the enzyme substrate binding site, the aromatic residues lining the active site play an important role; the sub sites are usually occupied by structural elements originating from the inhibitor molecule; the structure-segments and loop regions strongly involved in the inhibition process are likely to correspond to flexible components of the free structures of the molecules. Both mammalian and insect specific α-amylases dual inhibitors act through a common mechanism using alternative ways. Classification of Various alpha amylase inhibitors i. Involvement of lectin-like inhibitor from bean (Phaseolus vulgaris): (α -AI) [86,90,91] ii. A cereal-type α-amylase inhibitor: bifunctional ,α-amylase/trypsin inhibitor from ragi (Eleucine coracana Gaertneri; Indian Finger Millet) (RBI) [92] iii. A knottin-type α-amylase inhibitor, Amaranth α-amylase inhibitor (AAI) [81] iv. A Kunitz-like α-amylase inhibitor, Barley α -amylase/ subtilisin inhibitor (BASI) [93] v. Others Three-dimensional structure of insect Tenebrio molitor α-amylase enzyme (TMA) has been studied extensively. [40, 94]

B

Domain A is shown in blue, domain B in green, and domain C in red. The chloride anion and the calcium cation are represented by a purple and a yellow sphere, respectively, and the active-site residues Asp185, Glu222, and Asp287 are depicted in pink.(TMA, in common with almost all determined α-amylase structures (contains a calcium ion at a conserved position (yellow). The removal of the calcium ion in BLA causes local disorder around the Ca2+-binding site, resulting in an inactive enzyme. [95, 96] The three-dimensional model of insect a -amylases (TMA)comprises a single polypeptide chain of 471 amino-acid residues, one chloride ion, one calcium ion, and 261 water molecules. It displays elongated shape [7]. The protein folds into three distinct domains, named A, B and C (Fig. 1). The major structural unit of TMA is Domain A, composed of two segments (residues 1–97 and 160–379;represented as blue colour in Fig.1) Domain A contains the catalytic site and the ligand binding residues [7,95] Domain B is globular and is inserted into domain A. It is formed by several extended segments and a short α-helix (residue 98–159; green in Fig. 1). This domain forms a cavity against the ß?barrel of domain A in which the calcium ion is bound. Insect enzymes require one essential calcium ion to maintain their structural integrity, and are activated by chloride ions.[7, 93] Domain C is located exactly opposite to domain B on the other side of domain A. The C domain comprises the C-terminal residues 380–471 (red in Fig. 1) and forms a separated folding unit, exclusively made of ßsheet. Elements from domain A and B are involved in the architecture of the three most functionally important sites: the active site, the calcium-binding site, and the chloride-binding site [7,97-99]

Table 3.Common sources of amylase inhibitors and their effect on insect mortality Source of Insect Affected α-amylase inhibitor

A

Corms of Colocasia Carica papaya seeds CpAI Phaseolus vulgaris Phaseolus vulgaris WRP24 (T. aestivum) Phaseolus vulgaris (a AI-1) Wheat, 0.19 inhibitor Wheat α-amylase Inhibitor

Effect

Concentration

Ref.

C.cephalonica larvae 100% larval mortality Callosobruchus maculates 50% larval mortality 50% adult fecundity

0.0036% w/w 1.5% w/w 1.5% w/w

[35] [78]

AcanthoscelidesObtectus Zabrotes subfasciatus T.castaneum larvae

80–85% larval mortality 80–85% larval morality Weight loss 17.7 + 2.8 %

4% w/w 4% w:w 10% w/w

[36]

T.castaneum

larval weight

1% w/w

[34]

0.5% w/w 0.10% 1% 2% 1%

[33] [38]

Acanthoscelides Obtectus Larval weight loss 80% Rhyzopertha mortality 3.0 % dominica mortality 18.8% mortality 40.6 % weight loss 3.6 %

C

weight loss 21.9%

Figure 1 Ribbon plot of TMA. (fig. 1 taken from [92])

[37]

2%

i.Lectin like á-amylase inhibitors: Insect alpha amylase The bean P. vulgaris α-amylase inhibitor interacting with TMA The seeds of the common bean, P. vulgaris, contain two forms of α -

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 amylase inhibitor α -AI1 and α -AI2 [100], which differ in both their primary sequence and their inhibitory activity towards bruchids [9,101,102]. The overall architecture of the bean-inhibitor corresponds to a classical lectin fold (Bompard-Gilles (PDB ID-code, 1DHK). Interaction of a -AI1 with the insect a -amylase TMA To elucidate the inhibitory mechanism of these inhibitors, the structures of the common bean a-AI1 in complex with TMA [91] have been determined. Although TMA has the structure typical of α–amylases [92] (PDB ID-code, 1JAE). Structural analyses of the complex between a- AI1 and TMA [81] (PDB ID-code, 1viw) indicated that the strong contacts occurring between the inhibitor and the catalytic cleft of TMA enzymes are highly conserved. Structural analysis demonstrated that two hairpin loops of α-AI1 (residues 29–46 and 171–189) when inserted into the TMA reactive site it blocks substrate binding and establishes a hydrogen bond network with the residues of the substrate-binding region. The catalytic residues are strongly bonded to the inhibitor residues Tyr186 and Tyr37 that occupies the catalytic pocket. This class of inhibitors has been used for its insecticidal properties to protect seeds for insect predation [72, 73, 74]. ii.Cereal-type α-amylase inhibitors α-Amylase inhibitors of the cereal family are composed of 120–160 amino-acid residues forming five disulfide bonds [45,103]. A set of α -amylase inhibitors were purified from wheat flour,they display molecular masses of about 12, 24, and 60 kDa and are members of the cereal superfamily of α -amylase inhibitors. They act as monomers, homodimers or hetero-oligomers.The T. molitor α amylase (TMA) is effectively inhibited by a number of water-soluble protein components of the wheat kernel, particularly those termed inhibitors 0.28 and 0.19, which refers to their gel electrophoretic mobility relative to bromophenol blue [104]. The inhibitor 0.28 is a monomer with a molecular weight of 12 kDa, whereas the inhibitor 0.19 is a dimer with a molecular weight of about 24 kDa[105]. The dissociation constants of the a -amylase inhibitor 0.19 and α -amylase inhibitor 0.28 complexes are 0.85 and 0.13 nM, respectively [106]. The inhibitor referred to as 0.19 inhibits α-amylases from human saliva, pig pancreas, chick pancreas, the yellow mealworm, and Bacillus subtilis [45] The molecule has 124 amino acid residues [106] and its amino acid sequence is 26% identical to that of RBI [107]. It is the only other member of the cereal family with a known 3-D structure (PDB ID-code, 1HSS) [108] In the crystal structure, the asymmetric unit has four molecules of 0.19, each corresponding to a monomer of 124 amino acid residues. Each subunit consists of four major a-helices arranged in an ‘‘up and down’’ pattern and two short antiparallel h-strands [108]. As the structure of RBI–TMA complex reveals, the inhibitor binds to the enzyme active site, Two RBI segments are responsible for the interaction with TMA. Segment 1, comprising the N-terminal residues Ser1–Ala11 and the residues Pro52–Cys55, protrudes like an arrow head into TMA’s substrate-binding groove and

directly targets the active site of the enzyme. The Ser1 residue makes several hydrogen bonds with the catalytic Asp185 and Glu222 from enzyme while Val2 and Ser5 from inhibitor interact with the third conserved acidic residue of the catalytic site, Asp287. The second binding segment comprises several residues, which form a collar around the upper part of the arrow head and stabilize the complex. [92] iii.Knottin-type α-amylase inhibitors The amaranth α -amylase inhibitor AAI specifically inhibits a -amylases from insects, but not from mammalian sources [80] It is the smallest proteinaceous inhibitor of α-amylases with just 32 residues and three disulfide bonds .Based on its overall fold, its threestranded twisted sheet and the topology of its disulfide bonds, AAI has been classified as a knottin-like protein [109,110] Interaction of AAI with the insect α-amylase TMA The structural analysis of the AAI–TMA complex [81] provides high specificity of the AAI inhibitor against insect α–amylase, due to high complementarily of the interaction surfaces TMA–AAI complex.The crystal structure of TMA in complex with AAI was determined at 2.0 A ° resolution (PDB ID-code, 1CLV) [81]. AAI specifically inhibits insect α-amylases and is inactive against mammalian a-amylases [80] The structure of its inhibitor in complex revealed that inhibition, as with the lectin-like inhibitors, is through blockage of the catalytic site [81]. The inhibitor binds in the active site crevice interacting with catalytic residues from the A and B domains of TMA The residue Asp287, one of the catalytic residues of its enzyme, forms a salt bridge directly with Arg7 of AAI. The other two enzymatic catalytic residues, as well other conserved residues involved in substrate recognition and orientation, are connected to AAI via an intricate watermediated hydrogen-bonding network [81] iv.Kunitz-like α-amylase inhibitors The Kunitz-like α-amylase inhibitors contain around 180 residues and four cystines .They are present in cereals such as barley [111], wheat [112] and rice [113] The best-characterized α-amylase inhibitor from the Kunitz class is the barley α-amylase/subtilisin inhibitor (BASI), a bifunctional double-headed inhibitor with a fast tight inhibitory reaction with cereal α-amylase AMY2 (Ki = 0.22 nm) and serine proteinases of the subtilisin family [47] The structure of BASI [93] revealed two disulfide bonds and a ß-trefoil topology shared with the homologous wheat α-amylase subtilisin inhibitor (WASI) [114] the Erythrina caffra trypsin inhibitor [115] and the ricin B chain [116]. The mechanism of inhibition of barley α-amylase 2 (AMY2) by BASI is different, in that the inhibitor does not interact directly with any catalytic acidic residues of the enzyme. Nevertheless, this inhibitor interacts strongly with both the A and B domains near the catalytic site, through the formation of 12 hydrogen bonds, two salt bridges and multiple van der Waal’s contacts, and thereby prevents substrate access. A cavity at the enzyme–inhibitor interface contains a trapped calcium ion whose presence is suggested to electrostatically enhance the network of water molecules at the complex interface and thereby raises the stability of the complex.

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 v.Others classes of inhibitors toxicity and pathogenesis increase there is also the potential to identifying new ecologically safe means of plant defence , develop and Thaumatin-like α-amylase inhibitors type select safer bio pesticides having least effect on human population in The thaumatin-like inhibitors are proteins with molecular masses of the future. The review indicates that these enzyme inhibitors may =22 kDa with significant sequence similarity to pathogenesis-related play an active role in protecting plants from pests and causative group 5 (PR-5) proteins and to thaumatin, an intensively sweet pro- agents of diseases. tein from Thaumatococcus danielli [117] REFERENCES The best-characterized inhibitor from this class is zeamatin, a bifunc1. Oerke EC, Dehne HW: Safeguarding production – losses in tional inhibitor from Zea mays that is homologous to the sweet promajor crops and the role of crop protection, Crop Prot, 2004, 23, 275–285. tein thaumatin. Zeamatin was able to inhibit porcine pancreatic trypsin 2. Oerke EC: Crop losses to pests, J Agric Sci, 2006, 144,31–43. and digestive α-amylases of the insects T. castaneum, Sitophilus 3. Terra WR, Ferreira C: Insect digestive enzymes: properties, zeamays and Rizopherta dominica [89] compartmentalization and function, Comp Biochem Physiol, 1994, 109B,1–62. Zeamatin has a total of 13 ß-strands, 11 of which form a ß-sandwich at 4. Carlini CR, Grossi-de-Sá MF: Plant toxic proteins with insecthe core of protein [118]. Several loops extend from this inhibitor core ticidal properties. A review on their potentialities as and are secured by one or more of the eight disulfide bonds. Electrobioinsecticides, Toxicon, 2002, 40,1515–1539. static modelling of zeamatin reveals an electrostatically polarized sur5. Cinco-Moroyoqui FJ, Rosas-Burgos EC, Barboa-Flores J, face, heavily populated with Arg and Lys residues [118]. Other proCortez-Rocha MO: Alpha-amylase activity of Rhyzopertha teins from this class, such as the thaumatin-like proteins R and S from dominica (Coleoptera: Bostrichidae) reared on several wheat barley seeds, did not show any inhibitory activity against trypsin or varieties and its inhibition with kernel extracts, J Econ Entomol, 2006, 99,2146–2150. α-amylases [119] despite their highly similar N-terminal sequences. 6. Cinco-Moroyoqui FJ, Diaz-Malvaez FI, Alanis-Villa A, Zeamatin is mainly known for its antifungal activity binds to ß-1,3Barron-Hoyos JM, Cardenas-Lopez JL, Cortez-Rocha MO , glucans [120] and permeabilizes fungal-cells leading to cell death Wong-Corral, F.J.: Isolation and partial characterization of [121] It could be used as a medical agent, acting on vaginal murine three isoamylases of Rhyzopertha dominica F. (Coleoptera: candidosis cells [122] or in transgenic plants, increasing their resisBostrichidae), Comp Biochem. Physiol., 2008, 150B,153–160. tance against pests and pathogens. 7. Strobl S, Maskos K, Wiegand G, Huber R, Gomis-Ruth FX, Glockshuber R: A novel strategy for inhibition of alpha-amyγ-Purothionins-like α-amylase inhibitors lases: yel-low meal worm alpha-amylase in complex with the The α-amylase inhibitors of this family have 47 or 48 residues, are ragi bifunctional‘inhibitor at 2.5 A resolution, Structure, 1998, sulfur-rich and form part of the γ-thionin superfamily. Members of 6,911–921 this superfamily are involved in plant defence through a remarkable 8. Campos FAP, Xavier-Filho J, Silva CP, Amy MB: Resolution and partial characterization of proteinases and alpha amyvariety of mechanisms: modification of membrane permeability [123], lases from midgets of larvae of the bruchids beetle inhibition of protein synthesis [124] and proteinase inhibition [125] collasobruchus maculates (F.). Comp Biochem Physiol, 1989, Inhibition of insect α-amylases has been observed for three isoforms B 92, 51–57. from Sorghum bicolor called SIα-1, SIα-2 and SIα-3 [48]. These mol9. Grossi de Sa MF, Chrispeels MJ: Molecular cloning of ecules strongly inhibited the digestive α-amylases of guts of locust bruchid (Zabrotes subfasciatus) α-amylase cDNA and inand cockroach, poorly inhibited α-amylases from A. oryzae and huteractions of expressed enzyme with bean amylase inhibiman saliva and failed to inhibit the α-amylases from porcine pantors, Insect Biochem Mol Biol, 1997, 27,271–281. creas, barley and Bacillus sp. [48]The structure of SIα-1 revealed a 10. Aktar W., Sengupta D., Chawdhury A. Impact of pesticides α+ ß sandwich structure [126] with a nine-residue helix packed tightly use in agriculture: their benefits and hazards, Interdiscip against the sheet The helix is held in place by two disulfide bridges, Toxicol. 2009, 2(1): 1–12. which link sequential turns of the helix to residues 41 and 43 in the 11. Sharma H, Sharma K, Seetharama N, Ortiz R: Prospects for transgenic resistance to insects in crop improvement, Elect middle of strand ß3, the so-called cysteine-stabilized helix (CSH) moJ Biotechnol, 2000, 3,76-95 tif [127]. The structure is similar to those of wheat γ1-purothionin 12. World Resources Institute: World Resources, 1998/1999. [128] and scorpion toxins [127]. Oxford University Press, UK. 13. Bassil KL, Vakil C, Sanborn M, Cole DC, Kaur JS, Kerr KJ: CONCLUSION Cancer health effects of pesticides: systematic review, Can There is need for use of environment friendly yet efficient naturally Fam Physician, 2007, 53,1704–1711. occurring bio control agents for insects. The use of environmentally 14. Kanavouras K, Tzatzarakis MN, Mastorodemos V, Plaitakis benign insect pest control measures will be in demand only if it has A, Tsatsakis AM: A case report of motor neuron disease in a increased efficacy which can be achieved by incorporation of enpatient showing significant level of DDTs, HCHs and orga hancers, selective enzyme inhibitors and synergists into bio pestinophosphate metabolites in hair as well as levels of hexane cide formulations. As our knowledge of the molecular basis of

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458

15.

16. 17.

18.

19. 20.

21. 22.

23. 24. 25. 26.

27.

28.

29.

and toluene in blood, Toxicol Appl Pharmacol, 2011, 256,399– 404. Parrón T, Requena M, Hernández AF, Alarcón R: Association between environmental exposure to pesticides and neurodegenerative diseases, Toxicol Appl Pharmacol, 2011, 256,379–385. Hernández AF, Parrón T, Alarcón RB: Pesticides and asthma, Curr Opin Allergy Clin Immunol, 2011, 11,90–96. Hanke W, Jurewicz J: The risk of adverse reproductive and developmental disorders due to occupational pesticide exposure: an overview of current epidemiological evidence. Int J Occup Med Environ, 2004, 17,223–243. Pin˜a-Guzman B, Solý´s-Heredia MJ, Quintanilla-Vega B: Diazinon alters sperm chromatin structure in mice by phosphorylating nuclear protamines, Toxicology and Applied Pharmacology, 2005, 202,189– 198. Dalvie MA, London L: Risk assessment of pesticide residues in South African raw wheat, Crop Protection, 2009, 28,864–869. Guler GO, Cakmak YS, Dagli Z, Aktumsek A., Ozparlak H: Organochlorine pesticide residues in wheat from Konya region, Turkey, Food and Chemical Toxicology, 2010, 48,1218– 1221. Jaga K, Brosius D: Pesticide exposure: human cancers on the horizon, Reviews on Environmental Health, 1999, 14,3950. Ventura C, Núñez M, Miret N, Lamas DM, Randi A, Venturino A, Rivera E, Cocca C: Differential mechanisms of action are involved in chlorpyrifos effects in estrogen-dependent or independent breast cancer cells exposed to low or high concentrations of the pesticide, Toxicology Letters, 2012, 213,184–193. Darko G, Akoto O: Dietary intake of organophosphorus pesticide residues through vegetables from Kumasi, Ghana, Food and Chemical Toxicology, 2008,46,3703–3706. Chen C, Qian Y,Chen Q, Tao C, Li C, Yun Li: Evaluation of pesticide residues in fruits and vegetables from Xiamen, China, Food Control, 2011, 22,1114-1120. Farag AT, Okazy AM, El-Aswed AF: Developmental toxicity study of chlorpyrifos in rats, Reproductive Toxicology, 2003, 17, 203–208. Farag AT, Radwan AH, Sorour F, Okazy AE, El-Sayed ElAgamy, Abd El-Khaliek El-Sebae: Chlorpyrifos induced reproductive toxicity in male mice, Reproductive Toxicology, 2010, 29, 80–85. Lima CS, Nunes-Freitas AL, Ribeiro-Carvalho A, Filgueiras CC, Manhães AC, Meyer A, Abreu-Villaça Y: Exposure to methamidophos at adulthood adversely affects serotonergic biomarkers in the mouse brain, NeuroToxicology, 2011, 32,718–724. Maia LO, Júnior WD, Carvalho LS, Paiva GD, Araujo P, Costa MF, Andersen ML, Tufik S, Mazaro-Costa R: Association of methamidophos and sleep loss on reproductive toxicity of male mice, Environmental Toxicology and Pharmacology, 2011, 32,155–161. Spassova D, White T, Singh AK: Acute effects of acephate and methamidophos on acetylcholinesterase activity, endocrine system and amino acid concentrations in rats, Comparative Biochemistry and Physiology Part C: Pharmacol-

ogy, Toxicology and Endocrinology, 2000, 126,79–89. 30. Franco OL, Rigden DJ, Melo FR, Grossi-de-Sa MF: Plant αamylase inhibitors and their interaction with insect α-amylases: Structure, function and potential for crop protection, Eur J Biochem, 2002, 269, 397–412. 31. Andow DA: The risk of resistance evolution in insects to transgenic insecticidal crops, Collection of Biosafety Reviews,2008, 4,142–199 32. Sharma KK, Ortiz R: Program for the application of genetic transformation for crop improvement in the semi-arid tropics, In Vitro Cell Dev Biol Plant, 2000, 36, 83–92 33. Franco OL, Melo FR, Mender PA, Paes NS, Yokoyama M, Coutinho MV, Bloch C Jr, Grossi-de-sa MF Characterization of two Acanthoscelides obtectus α-amylase and their inactivation by wheat inhibitors. J Agric Food Chem, 2005, 53:1585-1590 34. Kluh I, Horn M, Hy´blova J, Hubert J, Lucie Dolec¡kova´Mares¡ova´ a, Zdene¡k Voburka a, Iva Kudlý´kova´ b, Frantis¡ek Kocourek b, Michael Mares: Inhibitory specificity and insecticidal selectivity of α-amylase inhibitor from Phaseolus vulgaris Phytochemistry, 2005, 66,31–39 35. Kumari B, Sharma P, Nath AK: α-Amylase inhibitor in local Himalyan collections of Colocasia: Isolation, purification, characterization and selectivity towards α-amylases from various sources, Pesticide Biochemistry and Physiology, 2012, 103,49–55 36. Gatehouse AMR, Dobie P, Hodges RJ, Meik J, Pusztai A, Boulter D: Role of carbohydrates in insect resistance in Phaseolus vulgaris, J Insect Physiol, 1987, 33,843–850. 37. Feng GH, Richardson M, Chen MS, Kramer KJ, Morgan TD, Reeck GR: Alpha-Amylase inhibitors from wheat: amino acid sequences and patterns of inhibition of insect and human alpha-amylases, Insect Biochem Mol Biol, 1996, 26, 419–426 38. Priya S, Kaur N, Gupta AK: Purification, characterization and inhibition studies of α-amylase of Rhyzopertha dominica. Pesticide Biochemistry and Physiology, 2010, 98, 231–237. 39. Marshall JJ, Lauda CM: Purification and properties of phaseolamin, an inhibitor of a-amvlase, from the kidney bean Phaseolus vulgaris, J Biol Chem, 1975, 250, 8030–8037. 40. Grossi-de-Sa MF, Mirkov TE, Ishimoto M, Colucci G, Bateman KS, Chrispeels MJ: Molecular characterization of a bean a-amylase inhibitor that inhibits the α-amylase of the Mexican bean weevil Zabrotes subfasciatus, Planta, 1997, 203,295–303. 41. Giri AP, Kachole MV: Amylase inhibitors of pigeonpea (Cajanus cajan) seeds, Phytochemistry, 1998, 47,197-202. 42. Melo FR, Sales MP, Silva LS, Franco OL, Bloch C Jr, Ary MB: α-Amylase from cowpea seeds, Prot Pept Lett, 1999, 6,387-392. 43. Ryan CA: Protease inhibitors in plants: genes for improving defense against insects and pathogens, Annu Rev Phytopath, 1990, 28,425–449. 44. Yamagata H, Kunimatsu K, Kamasaka H, Kuramoto T, Iwasaki T: Rice bifunctional α-amylase/subtilisin inhibitor: characterization, localization, and changes in developing and germinating seeds, Biosc Biotechnol Biochem, 1998, 62, 978– 985. 45. Franco OL, Ridgen DJ, Melo FR, Bloch Jr. C, Silva CP, Grosside-Sa MF: Activity of wheat α-amylase inhibitors towards

BioMedRx Vol.1 Issue 5 .May 2013

449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458

46.

47.

48.

49.

50.

51.

52.

53.

54.

55.

56.

57.

58.

bruchid α-amylase inhibitors and structural explanation of observed specificities, Eur J Biochem, 2000, 267,2166–2173 Petrucci T, Rab A, Tomasi M, Silano V: Further characterization studies of the alpha-amylase protein inhibitor of gel eletrophoretic mobility 0. 19 from the wheat kernel, Biochim Biophys Acta, 1976, 420, 288–297. Abe JI, Sidenius U, Svensson B: Arginine is essential for the α-amylase inhibitory activity of the α-amylase/subtilisin inhibitor (BASI) from barley seeds, Biochem J, 1993, 293,151– 155. Bloch Jr. C, Richardson M: A new family of small (5kD) protein inhibitors of insect α-amylase from seeds of sorghum (Sorghum bicolor (L.) Moench) have sequence homologies with wheat δ-purothionins, FEBS Lett, 1991, 279, 101–104. Iulek J , Franco OL, Silva M, Slivinski CT, Jr CB, Rigden DJ,Grossi de Sa´ MF: Purification, biochemical characterisation and partial primary structure of a new aamylase inhibitor from Secale cereale (rye), The International Journal of Biochemistry & Cell Biology, 2000, 32,1195– 1204. Garcia-Casado GL, Sanchez-Monge R, Lopez-Otin C, Salcedo G: Rye inhibitors of animal α-amylases shown different specificities, aggregative properties and IgE-binding capacities than their homologues from wheat and barley, Eur J Biochem, 1994, 224, 525–531. Young NM, Thibault P, Watson DC, Chrispeels MJ: Posttranslational processing of two α-amylase inhibitors and an arcelin from the common bean, Phaseolus vulgaris, FEBS Lett, 1999, 446, 203–206 Sharma KK, Pattabiraman TN: Natural plant enzyme inhibitors, Isolation and characterization of two α-amylase inhibitors from Colocasia antiquarum tubers, J Sci Food Agric, 1980, 31,981–991. McEwan R, Madivha RP, Djarova T, Oyedeji OA, Opoku AR: Alpha-amylase inhibitor of amadumbe (Colocasia esculenta): Isolation, purification and selectivity toward aamylases from various sources, Afr J Biochem, 2010, Res.4, 220–224. Shivaraj B, Sharma KK, Pattabiraman TN: Natural plant enzyme inhibitors.VIII. α-amylase inhibitors and amylases in plant tubersm, Indian J Biochem Biophys, 1979, 16, 52–55. Sharma KK, Pattabiraman TN: Natural plant enzyme inhibitors. Purification and properties of an amylase inhibitor from yam (Dioscorea alata), J Sci Food Agric, 1982, 33, 255–262. Glare T, Caradus J, Gelernter W, Jackson T, Keyhani N, Köhl J, Marrone P, Morin L, Stewart A: Have biopesticides come of age, Trends in Biotechnology, 2012, 30,250-258 Andrews RE, Faust RM, Wabiko H, Raymond KC, Bulla LA: The biotechnology of Bacillus thuringiensis, Crit Rev Biotech, 1987, 6,163–232. Vaeck M, Reynaerts A, Hofte H, Jansens S, de Beuckleer M, Dean C, Zabeau M, Van Montagu M, Leemans J: Transgenic

plants protected from insect attack, Nature, 1987, 328,33–37. 59. Vazquez-Padron RI, Moreno-Fierros L, Neri-Bazan L, de la Riva GA, Lopez-Revilla R: Intragastric and intraperitoneal administration of Cry1Ac protoxin from Bacillus thuringiensis induces systemic and mucosal antibody responses in mice, Life Sci, 1999, 64,1897–1912. 60. Vazquez-Padron RI, Gonzales-Cabrera J, Garcia-Tovar C, NeriBazan L, Lopez-Revilla R, Hernandez M, Moreno- Fierro L, de la Riva GA: Cry1Ac protoxin from Bacillus thuringiensis sp kurstaki HD73 binds to surface proteins in the mouse small intestine, Biochem Biophys Res Commun, 2000, 271,54– 58. 61. Zhu-Salzman K, Shade RE, Koiwa H, Salzman RA, Narasimhan M, Bressan RA, Hasegawa PM, Murdock LL: Carbohydrate binding and resistance to proteolysis control insecticidal activity of Griffonia simplicifolia lectin II, Proc Natl Acad Sci USA, 1998, 95,15123–15128. 62. Paes NS, Gerhardt IR, Coutinho MV, Yokoyama M, Santana E, Harris N, Chrispeels MJ, Grossi-de-Sa MF: The effect of arcelin-1 on the structure of the midgut of bruchid larvae and immunolocalization of the arcelin protein, J Insect Physiol, 2000, 46,393–402. 63. Ye X, Ng TB: A chitinase with antifungal activity from the mung bean, Protein Expr Purif, 2005, 40, 230–236. 64. Bishop JG, Ripoll DR, Bashir S, Damasceno CM, Seeds JD, Rose JK: Selection on glycine b-1,3-endoglucanase genes differentially inhibited by a Phytophthora glucanase inhibitor protein, Genetics, 2005, 169, 1009–1019. 65. Thevissen K, Idkowiak-Baldys J, Im YJ, Takemoto J, Francois IE, Ferket KK, Aerts AM, Meert EM, Winderickx J, Roosen J, Cammue BP: SKN1, a novel plant defensin-sensitivity gene in Saccharomyces cerevisiae, is implicated in sphingolipid biosynthesis, FEBS Lett, 2005, 579,1973–1977. 66. Chrispeels MJ, Grossi-de-Sa MF, Higgins TJV: Genetic engineering with alpha-amylase inhibitors makes seeds resistant to bruchids, Seed Sci Res, 1998, 8,257–263. 67. Gatehouse AMR, Gatehouse JA: Identifying proteins with insecticidal activity: use of encoding genes to produce insect-resistant transgenic crops, Pest Sci, 1998, 52,165–175. 68. Valencia A, Bustillo AE, Ossa GE, Chrispeels MJ: α- Amylases of the coffee berry borer (Hypothenemus hampei ) and their inhibition by two plant amylase inhibitors, Insect Biochem. Mol Biol, 2000, 30, 207–213. 69. Ishimoto M, Kitamura K: Growth inhibitory effects of an aamylase inhibitor from kidney bean, Phaseolus vulgaris (L.) on three species of bruchids (Coleoptera: Bruchidae), Appl Entomol Zool, 1989, 24,281–286. 70. Huesing JE, Shade RE, Chrispeels MJ, Murdock LL: α-Amylase inhibitor, not phytohemagglutinin explains the resistance of common bean seeds to cowpea weevil, Plant Physiol, 1991, 96,993–996. 71. Ishimoto M, Sato T, Chrispeels MJ, Kitamura K: Bruchid resistance of transgenic azuki bean expressing seed aamylase inhibitor of common bean, Entomol Exp Appl, 1996, 79,309– 315.

BioMedRx Vol.1 Issue 5 .May 2013

449-458

72.

73.

74.

75.

76.

77.

78.

79.

80.

81.

82.

83.

84.

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 Espindola Laila S.: Inhibitory action of Cerrado plants against Shade RE, Schroeder HE, Pueyo JJ, Tabe LM, Murdock LL, mammalian and insect α-amylases, Pesticide Biochemistry Higgins TJV, Chrispeels MJ: Transgenic pea seeds expressand Physiology, 2009, 95,141-146. ing the α-amylase inhibitor of the common bean are resis85. Titarenko E, Chrispeels MJ: cDNA cloning, bio- chemical tant to bruchid beetles, Bio Technology, 1994, 12,793–796. characterization and inhibition by plant inhibitors of the aSchroeder HE, Gollash S, Moore A, Tabe LM, Craig S, Hardie amylases of the Western corn rootworm, Diabrotica virgifera D, Chrispeels MJ, Spencer D, Higgins TJV: Bean α-amylase virgifera, Insect Biochem. Mol. Biol, 2000, 30,979-990. inhibitor confers resistance to the pea weevil, Bruchus 86. Nahoum V, Roux G, Anton V, Rouge P, Puigserver A, Bischo€ pisorum, in genetically engineered peas (Pisum sativum L.), H, Henrissat B, Payan F: Crystal structures of human panPlant Physiol, 1995, 107,1233–1239. creatic α-amylase in complex with carbohydrate and pro Morton RL, Schroeder HE, Bateman KS, Chrispeels MJ, teinaceous inhibitors, Biochem. J., 2000, 346,202-208. Armstrong E, Higgins TJV: Bean α-amylase inhibitor- I in 87. Silva CP, Terra WR, Xavier-Filho J, Grossi-de-Sa MF, transgenic peas (Pisum sativum ) provides complete protecIsejima EM, Damatta RA, Miguens FC, Bifano TD: Digestion from pea weevil (Bruchus pisorum ) under field condition of legume starch granules by larvae of Zabrotes tions, Proc Natl Acad Sci USA, 2000, 97,3820–3825. subfasciatus (Coleoptera: Bruchidae) and the induction of Pusztai A, Bardocz GG, Alonso R, Chrispeels MJ, Schroeder α-amylases in response to di€erent diets, Insect Biochem HE, Tabe LM, Higgins TJ: Expression of the insecticidal Mol Biol, 2001, 31, 41-50. bean alpha-amylase inhibitor transgene has minimal detri88. Sanchez-Monge R, Gomez L, Garcia-Olmedo F, Salcedo G: mental effect on the nutritional value of peas fed to rats at New dimeric inhibitor of heterologous α-amylases encoded 30% of the diet, J Nutr, 1999, 129,1597–1603. by a duplicated gene in the short arm of chromosome 3B of Francàoise P: Structural basis for the inhibition of mammawheat (Triticum aestivum L.), Eur J Biochem, 1989, 183,37lian and insect α-amylases by plant protein inhibitors, 40. Biochimica et Biophysica Acta, 2004, 696,1– 18. 89. Schimoler-O’Rourke R, Richardson Mand Selitrenniko CP: Pereira RA, Batista JAN, Mattar da Silva MS, Neto OB, Zeamatin inhibits trypsin and α-amylase activities, Appl Figueira ELZ, Jiménez AV, Grossi-de-Sa MF: An α-amylase Environ Microbiol, 2001, 67, 2365-2366. inhibitor gene from Phaseolus coccineus encodes a protein 90. Bompard-Gilles C, Rousseau P, Rouge P, Payan F: Substrate with potential for control of coffee berry borer mimicry in the active center of a mammalian alpha-amylase: (Hypothenemus hampei), Phytochemistry, 2006, 67,2009structural analysis of an enzyme–inhibitor complex, Struc2016 ture, 1996, 4,1441–1452. Farias LR, Costa FT, Souza LA, Pelegrini PB, Grossi-de-Sá 91. Nahoum V, Farisei F, Le-Berre-Anton V, Egloff MP, Rouge P, MF, Neto SM, Bloch Jr C, Laumann RA, Noronha EF, Franco Poerio E, Payan F: A plant-seed inhibitor of two classes of OL: Isolation of a novel Carica papaya α-amylase inhibitor alpha-amy-lases: X-ray analysis of Tenebriomolitorlarvae with deleterious activity toward Callosobruchus maculate,. alpha-amylase in complex with the bean Phaseolus vulgaris Pesticide Biochemistry and Physiology, 2007, 87,255–260 inhibitor, Acta Crystal-logr, 1999, D 55,360–362. Mehrabadi M, Bandani AR, Mehrabadi R, Alizadeh H: In92. Strobl S, Maskos K, Betz M, Weigand G, Huber R, Gomishibitory activity of proteinaceous α-amylase inhibitors from Ruth FX, Glockshuber R,: Crystal structure of yellow mealTriticale seeds against Eurygaster integriceps salivary aworm α-amylase at 1.64A resolution, J Mol Biol, 1998, amylases: Interaction of the inhibitors and the insect diges278,617–628. tive enzymes, Pesticide Biochemistry and Physiology, 2012, 93. Valle F, Kadziola A, Bourne Y, Juy M, Rodenburg KW, Svens102,220–228. son B, Haser R: Barley α-amylase bound to its endogenous Chagolla-LoÂpez A, Blanco-Labra A, Patthy A, SaÂnchez, protein inhibitor BASI: crystal structure of the complex at R, Pongor S: A novel α-amylase inhibitor from Amaranth 1.9 A resolution, Structure, 1998, 6,649–659. (Amaranthus hypocondriacus) seeds, J Biol Chem, 1994, 94. Titarenko E, Chrispeels MJ: cDNA cloning, bio- chemical 269, 23675-23680. characterization and inhibition by plant inhibitors of the αPereira PJB, Lozanov V, Patthy A, Huber R, Bode W, Pongor amylases of the Western corn rootworm, Diabrotica virgifera S, Strobl S: Speci®c inhibition of insect α-amylases: yellow virgifera, Insect Biochem. Mol. Biol., 2000, 30,979-990. meal worm α-amylase in complex with the Amaranth α-amy95. Hwang KY, Song HK, Chang C, Lee J, Lee SY, Kim KK, Choe lase inhibitor at 2.0 A Ê resolution: Structure,1999, 7,1079S, Sweet RM, Suh SW, Crystalstruc-ture of thermostable a1088. amylase from Bacillus licheniformisre®ned at 1.7 A Ê resoluBalnco-Wra A , Sandoval-Cardoso L, Mendiola-Olaya E, tion, Mol. Cells, 1997, 7,251-258. Valdes-Rodiuguez S, Lopez MG: Purification and Character96. Qian M , Haser R, Payan F: Structure and molecular model ization of a Glycoprotein α-Amylase Inhibitor from Tepary refine-ment of pig pancreatica-amylase at 2.1 A resolution, Bean Seeds (Phaseolus acutifolius A. Gray), J Plant Physiol, 1993, J Mol Biol 231,785–799. 1996, 149,650-656. 97. Brayer GD, Luo Y, Withers SG: The structure of human panYamada T, Hattori K, Ishimoto M: Purification and characcreatic α-amylase at 1.8 A° resolution and comparison with terization of two α-amylase inhibitors from seeds of tepary related enzymes, Protein Sci, 1995, 4, 1730–1742 bean (Phaseolus acutifolius A. Gray), Phytochemistry, 2001, 98. Ramasubbu N, Paloth V, Luo Y, Brayer GD, Levine MJ: Struc58,59–6 ture of human salivarya-amylase at 1.6 A °resolution: impliSilva Everton M., Valencia Arnubio, Grossi-de-Sá, Maria cations for its role in the oral cavity, ActaCrystallogr, 1996, Fátima, Rocha, Thales L. Freire, Érika, de Paula, José E., D52, 435–446 BioMedRx Vol.1 Issue 5 .May 2013 449-458

Neeraj Wadhwa et al. /BioMedRx 2013,1(5),449-458 teinase K/α-amylase inhibitor from wheat (PK13) at 2.5 A Ê 99. Chrispeels MJ, Raikhel NV: Lectins, lectin genes and their resolution, FEBS Lett, 1991, 279, 240-242. role in plant defence, Plant Cell, 1991, 3,1–9. 114. Onesti S, Brick P, Blow DM: Crystal structure of a Kunitz100. Hoffman LM, Donaldson DD: Characterization of two type trypsin inhibitor from Erythrinaca reseeds, J Mol Biol, Phaseolusvulgarisphytohaemagglutinin genes closely linked 1991, 217, 153-176. on the chromo-some, Embo J, 1985, 4, 883–889. 115. Rutenber E and Robertus JD: Structure of ricin B chain at 2.5 101. Suzuki K, Ishimoto M, Kitamura K: cDNA sequence and A resolution, Proteins, 1991, 10, 260-269. deduced primary structure of ana-amylase inhibitor from a 116. Vigers A, Roberts Wand Sellitrenniko CP: A new family of bruchid-resistant wild common bean, Biochim. Biophys Acta, plant antifungal proteins, Mol Plant Microb Interact, 1991, 1994, 1206, 289–291. 4, 315-323. 102. Lyons A, Richardson M, Tatham AS, Shewry PR: Character117. Batalia MA, Monzingo AF, Ernst S, Roberts W, Robertus ization of homologous inhibitors of trypsin and α-amylase, JD: The crystal structure of the antifungal protein zeamatin, Biochim Biophys Acta, 1987, 915, 305-313. amember of the thaumatin-like, PR-5 protein-family, Nat Struct 103. Silano V, Furia M, Gianfreda L, Macri A, Palescandolo R, Rab Biol, 1996, 3,19-23. A, Scardi V, Stella E, Valfre F: Inhibition of amylases from 118. Hejgaard J, Jacobsen S, Svendsen I: Two antifungal different origins by albumins from the wheat kernel, Biochim thaumatin-like proteins from barley grain, FEBS Lett, 1991, Biophys Acta, 1975, 391, 170–178. 291, 127-131. 104. Silano V, Pocchiari F, Kasarda DD: Physical characterization 119. Trudel J,Grenier J,Ptovin CandAsselin A: Several thaumatinof alpha-amylase inhibitors from wheat, Biochim Biophys like proteins bind tob-1,3-glucans, Plant Physiol, 1998, Acta, 533, 181–185. 118,1431-1438. 105. Maeda K, Kakabayashi S, Matsubura H: Complete amino 120. Abad LR, D’Urzo MP, Lin D, Narasimhan ML, Renveni M, acid sequence of α-amylase inhibitor in wheat kernel (0.19Zhu JK, Niu X, Singh NK, Hasegawa PM, Bressan RA: Anti inhibitor), Biochim Biophys Acta, 1995, 828, 213–221. fungal activity of tobacco osmotin has speci®city and in106. Campos FAP, Richardson M: The complete amino acid sevolves plasma membrane permeabilization, Plant Sci, 1996, quence of the bifunctional-amylase trypsin inhibitor from 118,11-23. seeds of ragi (India finger millet Eleusinecoracana, Gaertneri), 121. Selitrenniko CP,Wilson SJ, Clemons KV, StevensDA: FEBS Lett, 1983, 152, 300–304. Zeamatin, an antifungal protein, Curr Opin Anti-Infective 107. Oda Y, Matsunaga T, Fukuyiama K, Miyasaki T, Morimoto Drugs, 2000, 2, 368-374. JT: Tertiary and quaternary structures of 0.19-amylase in122. Castro MD, Fontes W, Morhy Land Bloch Jr C: Complete hibitor from wheat kernel determined by X-ray analysis at amino acide sequence from g-thionins from maize 2.06 A° resolution, Biochemistry, 1997, 36, 13503–13511. (ZeamaysL.) seeds, Protein Pept Lett, 1996, 3, 267-274. 108. Lu S, Deng P, Liu X, Luo J, Han R, Gu X, Liang S, Wang X, Li 123. Mendez E, Rocher A, Calero M, GirbesT, Citores Land F, Lozanov V, Patthy A, Pongor S: Solution structure of the Soriano F: Primary structure of x-hordothionin, a member of major α-amylase inhibitor of the crop plant amaranth, J Biol a novel family of thionins from barley endosperm, and its Chem, 1999, 274,20473-20478. inhibition of protein synthesis in eukaryotic and prokary109. Martins JC, Enassar M, Willen R, Wieruzeski JM, Lippens G, otic cell-free systems, Eur J Biochem, 1996, 239, 67-73. Wodak SJ: Solution structure of the main α-amylase inhibi124. Wijaya R, Neumann GM, Condron R, Hughes AB, PolyaGM: tor from amaranth seeds, Eur J Biochem, 2001, 268, 2379Defense proteins from seed ofCassia ®stula include a lipid 2389. transfer protein homologue and a protease inhibitory plant 110. Rodenburg KW, Varallyay E, SvendsenI and Svensson B: defensin, Plant Sci, 2000, 159,243-255. Arg-27, Arg-127 and Arg-155 in the b-trefoil protein barley 125. Orengo CA and Thornton JM: Alpha plus beta folds revisα-amylase/subtilisin inhibitor are interface residues in the ited: some favoured motifs, Structure1, 1993, 105-120. complex with barley a-amylase 2, Biochem J, 1995, 309, 969126. Bloch Jr C, Patel SU, Baud F, Zvelebil MJJM, Carr MD, Sadler 976. PJ , ThorntonJM: H-NMRstructure of an antifungal c-thionin 111. Gvozdeva EL, Valueva TA, Mosolov VV: Enzymatic oxidaprotein Sia1: similarity to scorpion toxins, Proteins, 1998, tion of the bifunctional wheat inhibitor of subtilisin and en32,334-349. dogenous α-amylase, FEBS Lett, 1993, 334, 72-74. 112. Ohtsubo K and Richardson M: The amino acid sequence of 127. Bruix M, JimeÂnz MA, Santoro J, Gonza Âlez C, Colilla FJ, a 20-kDa bifunctionalsubtilisin/α-amylase inhibitor from MeÂndez E, Rico M: Solution structure ofc1 H and c1-P grain of rice (Oryza sativa L.) seeds, FEBS Lett, 1992, 309, 68thionins from barley and wheat endosperm determined by 1 72. H-NMR: a structural motif common to toxic arthropod pro113. Zemke KJ, MuÈller-Fahrnow A, Jany KD, Pal GP, Saenger teins, Biochemistry, 1993, 32,715-724. W: The three-dimensional structure of the bifunctional pro

Source of support: Nil, Conflict of interest: None Declared

BioMedRx Vol.1 Issue 5 .May 2013

449-458