전체메뉴
검색
Article Search

JMB Journal of Microbiolog and Biotechnology

QR Code QR Code

Note

References

  1. Bao L, Huang Q, Chang L, Sun Q, Zhou J, Lu H. 2012. Cloning and characterization of two β-glucosidase/xylosidase enzymes from yak rumen metagenome. Appl. Biochem. Biotechnol. 166: 72-86.
    CrossRef
  2. Da Lage JL, Feller G, Jane ek Š. 2004. Horizontal gene transfer from Eukarya to bacteria and domain shuffling: the α-amylase model. Cell Mol. Life Sci. 61: 97-109.
    CrossRef
  3. Feng Y, Duan CJ, Pang H, Mo XC, Wu CF, Yu Y, et al. 2007. Cloning and identification of novel cellulase genes from uncultured microorganisms in rabbit cecum and characterization of the expressed cellulases. Appl. Microbiol. Biotechnol. 75:319-328.
    CrossRef
  4. Ferrer M, Golyshina OV, Chernikova TN, Khachane AN, Reyes-Duarte D, Santos VA, et al. 2005. Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora. Environ. Microbiol. 7: 1996-2010.
    CrossRef
  5. Ferrer M, Ghazi A, Beloqui A, Vieites JM, López-Cortés N, Marín-Navarro J, et al. 2012. Functional metagenomics unveils a multifunctional glycosyl hydrolase from the family 43 catalysing the breakdown of plant polymers in the calf rumen. PLoS One 7: e38134.
    CrossRef
  6. Gong X, Gruninger RJ, Qi M, Paterson L, Forster RJ, Teather RM, McAllister TA. 2012. Cloning and identification of novel hydrolase genes from a dairy cow rumen metagenomic library and characterization of a cellulase gene. BMC Res. Notes 5: 566.
    CrossRef
  7. Guo H, Feng Y, Mo X, Duan C, Tang J, Feng J. 2008. Cloning and expression of a beta-glucosidase gene umcel3G from metagenome of buffalo rumen and characterization of the translated product. Sheng Wu G ong C heng X ue B ao (Chinese) 24: 232-238.
  8. Hostinová E, Jane ek Š, Gašper k J. 2010. Gene sequence, bioinformatics and enzymatic characterization of α-amylase from Saccharomycopsis fibuligera KZ. Protein J. 29: 355-364.
    CrossRef
  9. Janecek S. 2002. How many conserved sequence regions are there in the α-amylase family? Biologia 57(Suppl. 11): 29-41.
  10. MacGregor EA, Janecek S, Svensson B. 2001. Relationship of sequence and structure to specificity in the alpha-amylase family of enzymes. Biochim. Biophys. Acta 1546: 1-20.
    CrossRef
  11. Machovi M, Jane ek Š. 2003. The invariant residues in the α-amylase family: just the catalytic triad. Biologia (Bratisl) 58:1127-1132.
  12. Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 32: 426-428.
    CrossRef
  13. Morita H, Kuwahara T, Ohshima K, Sasamoto H, Itoh K, Hattori M, et al. 2007. An improved DNA isolation method for metagenomic analysis of the microbial flora of the human intestine. Microb. Environ. 22: 214-222.
    CrossRef
  14. Nimchua T, Thongaram T, Uengwetwanit T, Pongpattanakitshote S, Eurwilaichitr L. 2012. Metagenomic analysis of novel lignocellulose-degrading enzymes from higher termite guts inhabiting microbes. J. Microbiol. Biotechnol. 22: 462-469.
    CrossRef
  15. Rashamuse KJ, Visser DF, Hennessy F, Kemp J, Roux-van der Merwe MP, Badenhorst J, et al. 2013. Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library. Curr. Microbiol. 66: 145151.
    CrossRef
  16. Stam MR, Danchin EGJ, Rancurel C, Coutinho PM, Henrissat B. 2006. Dividing the large glycoside hydrolase family 13 into subfamilies: towards improved functional annotations of α-amylase-related proteins. Protein Eng. Des. Sel. 19: 555-562.
    CrossRef
  17. Streit WR, Schmitz RA. 2004. Metagenomics: the key to the uncultured microbes. Curr. Opin. Biotechnol. 7: 492-498.
  18. Tas s e L, Bercovici J , Pizzut-Serin S , Robe P, Tap J , Klopp C, et al. 2010. Functional metagenomics to mine the human gut microbiome for dietary fiber catabolic enzymes. Genome Res. 20: 1605-1612.
  19. Van der Kaaij RM, Jane ek Š, van der Maarel MJEC, Dijkhuizen L. 2007. Phylogenetic and biochemical characterization of a novel cluster of intracellular fungal α-amylase enzymes. Microbiology 153: 4003-4015.
    CrossRef
  20. Walter J, Mangold M, Tannock GW. 2005. Construction, analysis, and β-glucanase screening of a bacterial artificial chromosome library from the large-bowel microbiota of mice. Appl. Environ. Microbiol. 71: 2347-2354.
    CrossRef
  21. Warnecke F, Luginbühl P, Ivanova N, Ghassemian M, Richardson TH, Stege JT, et al. 2007. Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature 450: 560-565.
    CrossRef
  22. Xu B, Xu W, Yang F, Li J, Yang Y, Tang X, et al. 2013. Metagenomic analysis of the pygmy loris fecal microbiome reveals unique functional capacity related to metabolism of aromatic compounds. PLoS One 8: e56565.
    CrossRef
  23. Zhao S, Wang J, Liu K, Zhu Y, Bu D, Li D, Yu P. 2009. Screening and characterization of lipase from a metagenome library of dairy rumen microflora. Sheng Wu Gong Cheng Xue Bao (Chinese) 25: 869-874.
  24. Zhao S, Wang J, Bu D, Liu K, Zhu Y, Dong Z, Yu Z. 2010. Novel glycoside hydrolases identified by screening a Chinese Holstein dairy cow rumen-derived metagenome library. Appl. Environ. Microbiol. 76: 6701-6705.
    CrossRef
  25. Zhou J, Dong Y, Li J, Zhang R, Tang X, Mu Y, et al. 2012. Cloning, heterologous expression, and characterization of novel protease-resistant α-galactosidase from new Sphingomonas strain. J. Microbiol. Biotechnol. 22: 1532-1539.
    CrossRef

Related articles in JMB

More Related Articles

Article

Note

J. Microbiol. Biotechnol. 2014; 24(4): 447-452

Published online April 28, 2014 https://doi.org/10.4014/jmb.1310.10121

Copyright © The Korean Society for Microbiology and Biotechnology.

Cloning and Characterization of a Novel α-Amylase from a Fecal Microbial Metagenome

Bo Xu 1, 2, 3, Fuya Yang 1, Caiyun Xiong 1, Junjun Li 1, 2, 3, Xianghua Tang 1, 2, 3, Junpei Zhou 1, 2, 3, Zhenrong Xie 1, Junmei Ding 1, 2, 3, Yunjuan Yang 1, 2, 3 and Zunxi Huang 1, 2, 3*

1School of Life Science, Yunnan Normal University, Kunming 650500, China, 2Engineering Research Center of Sustainable Development and Utilization of Biomass Energy, Ministry of Education, Kunming 650500, China, 3Key Laboratory of Yunnan for Biomass Energy and Biotechnology of Environment, Kunming 650500, China, 4Key Laboratory of Enzyme Engineering, Yunnan Normal University, Kunming 650500, China

Received: November 5, 2013; Accepted: January 2, 2014

Abstract

To isolate novel and useful microbial enzymes from uncultured gastrointestinal
microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed.
The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed
amylolytic activity. Subcloning and sequence analysis of a positive clone led to the
identification a novel gene (amyPL) coding for α-amylase. AmyPL was expressed in Escherichia
coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the
first to report the molecular and biochemical characterization of a novel α-amylase from a
gastrointestinal metagenomic library.

Keywords: amylase, metagenome, function-based screening, gastrointestinal microorganisms, cloning, characterization

References

  1. Bao L, Huang Q, Chang L, Sun Q, Zhou J, Lu H. 2012. Cloning and characterization of two β-glucosidase/xylosidase enzymes from yak rumen metagenome. Appl. Biochem. Biotechnol. 166: 72-86.
    CrossRef
  2. Da Lage JL, Feller G, Jane ek Š. 2004. Horizontal gene transfer from Eukarya to bacteria and domain shuffling: the α-amylase model. Cell Mol. Life Sci. 61: 97-109.
    CrossRef
  3. Feng Y, Duan CJ, Pang H, Mo XC, Wu CF, Yu Y, et al. 2007. Cloning and identification of novel cellulase genes from uncultured microorganisms in rabbit cecum and characterization of the expressed cellulases. Appl. Microbiol. Biotechnol. 75:319-328.
    CrossRef
  4. Ferrer M, Golyshina OV, Chernikova TN, Khachane AN, Reyes-Duarte D, Santos VA, et al. 2005. Novel hydrolase diversity retrieved from a metagenome library of bovine rumen microflora. Environ. Microbiol. 7: 1996-2010.
    CrossRef
  5. Ferrer M, Ghazi A, Beloqui A, Vieites JM, López-Cortés N, Marín-Navarro J, et al. 2012. Functional metagenomics unveils a multifunctional glycosyl hydrolase from the family 43 catalysing the breakdown of plant polymers in the calf rumen. PLoS One 7: e38134.
    CrossRef
  6. Gong X, Gruninger RJ, Qi M, Paterson L, Forster RJ, Teather RM, McAllister TA. 2012. Cloning and identification of novel hydrolase genes from a dairy cow rumen metagenomic library and characterization of a cellulase gene. BMC Res. Notes 5: 566.
    CrossRef
  7. Guo H, Feng Y, Mo X, Duan C, Tang J, Feng J. 2008. Cloning and expression of a beta-glucosidase gene umcel3G from metagenome of buffalo rumen and characterization of the translated product. Sheng Wu G ong C heng X ue B ao (Chinese) 24: 232-238.
  8. Hostinová E, Jane ek Š, Gašper k J. 2010. Gene sequence, bioinformatics and enzymatic characterization of α-amylase from Saccharomycopsis fibuligera KZ. Protein J. 29: 355-364.
    CrossRef
  9. Janecek S. 2002. How many conserved sequence regions are there in the α-amylase family? Biologia 57(Suppl. 11): 29-41.
  10. MacGregor EA, Janecek S, Svensson B. 2001. Relationship of sequence and structure to specificity in the alpha-amylase family of enzymes. Biochim. Biophys. Acta 1546: 1-20.
    CrossRef
  11. Machovi M, Jane ek Š. 2003. The invariant residues in the α-amylase family: just the catalytic triad. Biologia (Bratisl) 58:1127-1132.
  12. Miller GL. 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 32: 426-428.
    CrossRef
  13. Morita H, Kuwahara T, Ohshima K, Sasamoto H, Itoh K, Hattori M, et al. 2007. An improved DNA isolation method for metagenomic analysis of the microbial flora of the human intestine. Microb. Environ. 22: 214-222.
    CrossRef
  14. Nimchua T, Thongaram T, Uengwetwanit T, Pongpattanakitshote S, Eurwilaichitr L. 2012. Metagenomic analysis of novel lignocellulose-degrading enzymes from higher termite guts inhabiting microbes. J. Microbiol. Biotechnol. 22: 462-469.
    CrossRef
  15. Rashamuse KJ, Visser DF, Hennessy F, Kemp J, Roux-van der Merwe MP, Badenhorst J, et al. 2013. Characterisation of two bifunctional cellulase-xylanase enzymes isolated from a bovine rumen metagenome library. Curr. Microbiol. 66: 145151.
    CrossRef
  16. Stam MR, Danchin EGJ, Rancurel C, Coutinho PM, Henrissat B. 2006. Dividing the large glycoside hydrolase family 13 into subfamilies: towards improved functional annotations of α-amylase-related proteins. Protein Eng. Des. Sel. 19: 555-562.
    CrossRef
  17. Streit WR, Schmitz RA. 2004. Metagenomics: the key to the uncultured microbes. Curr. Opin. Biotechnol. 7: 492-498.
  18. Tas s e L, Bercovici J , Pizzut-Serin S , Robe P, Tap J , Klopp C, et al. 2010. Functional metagenomics to mine the human gut microbiome for dietary fiber catabolic enzymes. Genome Res. 20: 1605-1612.
  19. Van der Kaaij RM, Jane ek Š, van der Maarel MJEC, Dijkhuizen L. 2007. Phylogenetic and biochemical characterization of a novel cluster of intracellular fungal α-amylase enzymes. Microbiology 153: 4003-4015.
    CrossRef
  20. Walter J, Mangold M, Tannock GW. 2005. Construction, analysis, and β-glucanase screening of a bacterial artificial chromosome library from the large-bowel microbiota of mice. Appl. Environ. Microbiol. 71: 2347-2354.
    CrossRef
  21. Warnecke F, Luginbühl P, Ivanova N, Ghassemian M, Richardson TH, Stege JT, et al. 2007. Metagenomic and functional analysis of hindgut microbiota of a wood-feeding higher termite. Nature 450: 560-565.
    CrossRef
  22. Xu B, Xu W, Yang F, Li J, Yang Y, Tang X, et al. 2013. Metagenomic analysis of the pygmy loris fecal microbiome reveals unique functional capacity related to metabolism of aromatic compounds. PLoS One 8: e56565.
    CrossRef
  23. Zhao S, Wang J, Liu K, Zhu Y, Bu D, Li D, Yu P. 2009. Screening and characterization of lipase from a metagenome library of dairy rumen microflora. Sheng Wu Gong Cheng Xue Bao (Chinese) 25: 869-874.
  24. Zhao S, Wang J, Bu D, Liu K, Zhu Y, Dong Z, Yu Z. 2010. Novel glycoside hydrolases identified by screening a Chinese Holstein dairy cow rumen-derived metagenome library. Appl. Environ. Microbiol. 76: 6701-6705.
    CrossRef
  25. Zhou J, Dong Y, Li J, Zhang R, Tang X, Mu Y, et al. 2012. Cloning, heterologous expression, and characterization of novel protease-resistant α-galactosidase from new Sphingomonas strain. J. Microbiol. Biotechnol. 22: 1532-1539.
    CrossRef