Bifunctional recombinant cellulase–xylanase (rBhcell-xyl) from the polyextremophilic bacterium Bacillus halodurans TSLV1 and its utility in valorization of renewable agro-residues

Springer Science and Business Media LLC - Tập 20 - Trang 831-842 - 2016
Gurdeep Rattu1, Swati Joshi1, T. Satyanarayana1
1 Department of Microbiology, University of Delhi South Campus, New Delhi, India

Tóm tắt

The thermostable bifunctional CMCase and xylanase encoding gene (rBhcell-xyl) from Bacillus halodurans TSLV1 has been expressed in Escherichia coli. The recombinant E. coli produced rBhcell-xyl (CMCase 2272 and 910 U L−1 xylanase). The rBhcell-xyl is a ~62-kDa monomeric protein with temperature and pH optima of 60 °C and 6.0 with T1/2 of 7.0 and 3.5 h at 80 °C for CMCase and xylanase, respectively. The apparent K m values (CMC and Birchwood xylan) are 3.8 and 3.2 mg mL−1. The catalytic efficiency (k cat/K m ) values of xylanase and CMCase are 657 and 171 mL mg−1 min−1, respectively. End-product analysis confirmed that rBhcell-xyl is a unique endo-acting enzyme with exoglucanase activity. The rBhcell-xyl is a GH5 family enzyme possessing single catalytic module and carbohydrate binding module. The action of rBhcell-xyl on corn cobs and wheat bran liberated reducing sugars, which can be fermented to bioethanol and fine biochemicals.

Tài liệu tham khảo

Adlakha N, Rajagopal R, Kumar S, Reddy VS, Yazdani SS (2011) Synthesis and characterization of chimeric proteins based on cellulase and xylanase from an insect gut bacterium. Appl Environ Microbiol l77:4859–4866 Alvarez TM, Paiva JH, Ruiz DM, Cairo JPLF, Pereira IO, Paixao DA, de Almeida RF et al (2013) Structure and function of a novel cellulase 5 from sugarcane soil metagenome. PLoS One 8(12):e83635 Archana A, Satyanarayana T (2003) Purification and characterization of a cellulase free xylanase of a moderate thermophile Bacillus licheniformis A99. World J Microbiol Biotechnol 9:53–57 Aspeborg H, Coutinho PM, Wang Y, Brumer H, Henrissat B (2012) Evolution, substrate specificity and subfamily classification of glycoside hydrolase family 5 (GH5). BMC Evol Biol 12:186 Bodelon G, Palomino C, Fernandez LA (2013) Immunoglobulin domains in Escherichia coli and other enterobacteria: from pathogenesis to applications in antibody technologies. FEMS Microbiol Rev 37:204–250 Bourgault R, Oakley AJ, Bewley JD, Wilce MC (2005) Three-dimensional structure of (1,4)-beta-d-mannan mannanohydrolase from tomato fruit. Protein Sci 14:1233–1241 Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding 1976. Anal Biochem 72:248–254 Collin T, Gerday C, Feller G (2005) Xylanase, xylanase families and extremophilic xylanase. FEMS Microbiol Rev 29:3–23 Fan Z, Werkman JR, Yuan L (2009) Engineering of a multifunctional hemicellulase. Biotechnol Lett 31:751–757 Felsenstein J (1985) Confidence limits on phylogenies: an approach using the Bootstrap. Evolution 39:783–791 Galbe M, Zacchi G (2012) Pretreatment: the key to efficient utilization of lignocellulosic materials. Biomass Bioenerg 46:70–78 Gao Z, Ruan L, Chen X, Zhang Y, Xu X (2010) A novel salt-tolerant endo-β-1,4-glucanase Cel5A in Vibrio sp. G21 isolated from mangrove soil. Appl Microbiol Biotechnol 87:1373–1382 Gaur R, Tiwari S (2015) Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07. BMC Biotechnol 15:19 Ghatge SS, Telke AA, Kang SH, Arulalapperumal V, Lee KW, Govindwar SP, Oh YBD et al (2014) Characterization of modular bifunctional processive endoglucanase Cel5 from Hahella chejuensis KCTC 2396. Appl Microbiol Biotechnol 98:4421–4435 Han L, Feng J, Zhang S, Ma Z, Wang Y, Zhang X (2012) Alkali pretreated of wheat straw and its enzymatic hydrolysis. Braz J Microbiol 43:53–61 Hansen SA (1975) TLC method for identification of mono, di and trisaccharides. J Chromatogr 107:224–226 Hong Y (2009) Cloning of the Bacillus subtilis AMX-4 xylanase gene and characterization of the gene product. J Microbiol Biotechnol 19:1514–1519 Juturu V, Wu JC (2014) Microbial cellulases: engineering, production and applications. Renew Sust Energ Rev 33:188–203 Kumar V, Satyanarayana T (2011) Applicability of thermo-alkali-stable and cellulase-free xylanase from novel thermo-halo-alkaliphilic Bacillus halodurans in producing xylooligosaccharides. Biotechnol Lett 33:2279–2285 Kumar V, Satyanarayana T (2013) Biochemical and thermodynamic characteristics of thermo-alkali-stable xylanase from a novel polyextremophilic Bacillus halodurans TSEV1. Extremophiles 17:797–808 Li X, Lin W, Gao P, Chen F (1998) Endoglucanase S, a novel endocellulase exhibiting exoglucanase activity from a newly isolated Streptomyces sp. LX. J Appl Microbiol 85:347–356 Liu J, Liu WD, Zhao XL, Shen WJ, Cao H, Cui ZL (2011) Cloning and functional characterization of a novel endo-beta-1,4-glucanase gene from a soil-derived metagenomic library. Appl Microbiol Biotechnol 89:1083–1092 Lynd LR, Weimer PJ, van Zyl WH, Pretorius IS (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66:506–577 Mawadza C, Hatti-Kaul R, Zvauya R, Mattiasson B (2000) Purification and characterization of cellulases produced by two Bacillus strains. J Biotechnol 83:177–187 Medve J, Karlsson J, Lee D, Tjerneld F (1998) Hydrolysis of microcrystalline cellulose by cellobiohydrolase I and endoglucanase II from Trichoderma reesei: adsorption, sugar production pattern, and synergism of the enzymes. Biotechnol Bioeng 59:621–634 Morais S, Barak Y, Lamed R, Wilson DB, Xu Q, Himme ME, Bayer EA (2012) Paradigmatic status of an endo- and exoglucanase and its effect on crystalline cellulose degradation. Biotechnol Biofuels 5:78 Naika GS, Kaul P, Prakash V (2007) Purification and characterization of a new endoglucanase from Aspergillus aculeatus. J Agric Food Chem 55:7566–7572 Ozaki K, Hayashi Y, Sumitomo N, Kawai S, Ito S (1995) Construction, purification, and properties of a truncated alkaline endoglucanase from Bacillus sp. KSM-635. Biosci Biotechnol Biochem 59:1613–1618 Park JI, Kent MS, Datta S, Holmes BM, Huang Z, Simmons BA, Sale KL et al (2011) Enzymatic hydrolysis of cellulose by the cellobiohydrolase domain of CelB from the hyperthermophilic bacterium Caldicellulosiruptor saccharolyticus. Bioresour Technol 102:5988–5994 Perez-Avalos O, Sanchez-Herrera LM, Salgado LM, Ponce-Noyola T (2008) A bifunctional endoglucanase/endoxylanase from Cellulomonas flavigena with potential use in industrial processes at different pH. Curr Microbiol 57:39–44 Poidevin L, Feliu J, Doan A, Berrin JG, Bey M, Coutinho PM et al (2013) Insights into exo- and endoglucanase activities of family 6 Glycoside Hydrolases from Podospora anserina. Appl Environ Microbiol 79:4220–4229 Rosano G, Ceccarelli EA (2014) Recombinant protein expression in Escherichia coli: advances and challenges. Front Microbiol 5:172 Santos CR, Paiva JH, Sforça ML, Neves JL, Navarro RZ, Cota J, Akao PK et al (2012) Dissecting structure-function-stability relationships of a thermostable GH5-CBM3 cellulase from Bacillus subtilis 168. Biochem J 441:95–104 Shi P, Tian J, Yuan T, Liu X, Huang H, Bai Y, Yang P et al (2010) Paenibacillus sp. strain E18 bifunctional xylanase–glucanase with a single catalytic domain. Appl Environ Microbiol 76:3620–3624 St. John FJ, Rice JD, Preston JF (2006) Characterization of XynC from Bacillus subtilis subsp. subtilis strain 168 and analysis of its role in depolymerization of glucuronoxylan. J Bacteriol 188:8617–8626 Tabka MG, Herpoel-Gimbert I, Monod F, Asther M, Sigoillot JC (2006) Enzymatic saccharification of wheat straw for bioethanol production by a combined cellulase xylanase and feruloyl esterase treatment. Enzyme Microb Tech 39:897–902 Teeri TT (1997) Crystalline cellulose degradation: new insight into the function of cellobiohydrolases. Trends Biotechnol 15:160–167 Uday USP, Choudhury P, Bandyopadhyay TK, Bhunia B (2016) Classification, mode of action and production strategy of xylanase and its application for biofuel production from water hyacinth. Int J Biol Macromol 82:1041–1054 Vyas NK (1991) Atomic features of protein–carbohydrate interactions. Curr Opin Struct Biol 1:723–740 Wang H, Squina F, Segato F, Mort A, Lee D, Pappan K et al (2011) High-temperature enzymatic breakdown of cellulose. Appl Environ Microbiol 2011:5199–5206 Yu HY, Li X (2015) Alkali-stable cellulase from a halophilic isolate, Gracilibacillus sp. SK1 and its application in lignocellulosic saccharification for ethanol production. Biomass Bioenerg 81:19–25 Yuan SF, Wu TH, Lee HL, Hsieh HY, Lin WL, Yang B, Chang CK et al (2015) Biochemical characterization and structural analysis of a bifunctional cellulase/xylanase from Clostridium thermocellum. J Biol Chem 290:5739–5748 Zaks A, Klibanov AM (1988) Enzymatic catalysis in nonaqueous solvents. J Biol Chem 263:3194–3201