AER  Vol.3 No.3 , September 2015
A Novel Exo-Glucanase Explored from a Meyerozyma sp. Fungal Strain
ABSTRACT
Isolating cellulase-secreting microbes followed-by screening their cellulolytic activities has been an essential approach to discover novel and potential cellulases for cellulolytic industrial applications. This study was aimed to explore competitive exoglucanases by screening avicelase activities for 92 fungal strains isolated from environmental airborne-fungal-spore samples. Results showed that an isolated fungal strain numbered 58 exhibited the best avicelase activity of 0.209 U/mL when cultured for six days at pH 5.0 - 5.3 and 25℃ - 27℃, and was lately identified as a yeast strain of Meyerozyma sp. (96% ITS fragment similar with Meyerozyma caribbica, HG970748). Based on amino acid sequences revealed from LC/MS/MS, the target exoglucanase was identical to 1,4-beta-D-glucan cellobiohydrolases and was named Mc-CBHI which had optimal avicelase reaction conditions of pH 5 and 70℃ and could remain fairly stable after 4hr incubation at acid conditions (pH 3 - 5) or wide temperature ranges (30℃ - 80℃). Additionally, the Mc-CBHI (~70 kDa and ~3.6% of crude enzyme) had specific FPase and avicelase activities of 0.179 U/mg and 0.126 U/mg, respectively (which were about 40% - 50% activities of a commercial cellulase Accellerase-1000). These results demonstrated that the newly-found Mc-CBHI could become one of potential exoglucanase resources for related cellulolytic industrial applications.

Cite this paper
Kuo, H. , Zeng, J. , Wang, P. and Chen, W. (2015) A Novel Exo-Glucanase Explored from a Meyerozyma sp. Fungal Strain. Advances in Enzyme Research, 3, 53-65. doi: 10.4236/aer.2015.33006.
References
[1]   Tian, C., Zheng, L., Miao, Q., Cao, C. and Ni, Y. (2014) Improving the Reactivity of Kraft-Based Dissolving Pulp for Viscose Rayon Production by Mechanical Treatments. Cellulose, 21, 3647-3654.
http://dx.doi.org/10.1007/s10570-014-0332-1

[2]   Rubin, E.M. (2008) Genomics of Cellulosic Biofuels. Nature, 454, 841-845.
http://dx.doi.org/10.1038/nature07190

[3]   Kuhad, R.C., Gupta, R. and Singh, A. (2011) Microbial Cellulases and Their Industrial Applications. Enzyme Research, 2011, Article ID: 280696.
http://dx.doi.org/10.4061/2011/280696

[4]   Lynd, L.R., Weimer, P.J., van Zyl, W.H. and Pretorius, I.S. (2002) Microbial Cellulose Utilization: Fundamentals and Biotechnology. Microbiology and Molecular Biology Reviews, 66, 506-577.
http://dx.doi.org/10.1128/MMBR.66.3.506-577.2002

[5]   Kaur, B., Cberoi, H.S. and Chadha, B.S. (2014) Enhanced Cellulase Producing Mutants Developed from Heterokaryotic Aspergillus Strain. Bioresource Technology, 156, 100-107.
http://dx.doi.org/10.1016/j.biortech.2014.01.016

[6]   Saini, R., Saini, J.K., Adsul, M., Patel, A.K., Mathur, A., Tuli, D. and Singhania, R.R. (2015) Enhance Cellulase Production by Penicillium oxalicum for Bio-Ethanol Application. Bioresource Technology, 188, 240-246.
http://dx.doi.org/10.1016/j.biortech.2015.01.048

[7]   Jahangeer, S., Khan, N., Sohail, M., Jahangeer, S., Sohail, M., Shahzad, S., Ahmad, A. and Khan, S.A. (2005) Screening and Characterization of Fungal Cellulases Isolated from the Native Environmental Source. Pakistan Journal of Botany, 37, 739-748.

[8]   Dar, R.A., Saba, I., Shahnawaz, M., Sangale, M.K., Ade, A.B., Rather, S.A. and Qazi, P.H. (2013) Isolation, Purification and Characterization of Carboxymethyl Cellulase (CMCase) from Endophytic Fusarium oxysporum Producing Podophyllotoxin. Advances in Enzyme Research, 1, 91-96.
http://dx.doi.org/10.4236/aer.2013.14010

[9]   Boyce, A. and Walsh, G. (2015) Characterisation of a Novel Thermostable Endoglucanase from Alicyclobacillus vulcanalis of Potential Application in Bioethanol Production. Applied Microbiology and Biotechnology.
http://dx.doi.org/10.1007/s00253-015-6474-8

[10]   Sapra, R., Park, J.I., Dalta, S. and Simmons, B.A. (2011) Novel Thermophilic Cellobiohydrolases. US Patent No. 20110207182 A1.

[11]   Brevnova, E.E. (2013) Isolation and Characterization of Schizochytrium aggregatum Cellobiohydrolases I (Cbh 1). US Patent No. 8470592 B2.

[12]   Yu, S.-M., Ho, T.H.D., Kuo, H.-W., Wu, N.I.S., Li, C.-W. and Ju, Y.-M. (2013) Beta-Glucosidase and Uses Thereof. US Patent No. 8394619 B1.

[13]   Kausar, H., Sariah, M., Saud, H.M., Alam, M.Z. and Ismail, M.R. (2010) Development of Compatible Lignocellulolytic Fungal Consortium of Rapid Composting of Rice Straw. International Biodeterioration & Biodegradation, 64, 594-600.
http://dx.doi.org/10.1016/j.ibiod.2010.06.012

[14]   Khokhar, I., Haider, M.S., Mushtaq, S. and Mukhtar, I. (2012) Isolation and Screening of Highly Cellulolytic Filamentous Fungi. Journal of Applied Sciences and Environmental Management, 16, 223-226.

[15]   Ja’afaru, M.I. (2013) Screening of Fungi Isolated from Environmental Samples for Xylanase and Cellulase Production. ISRN Microbiology, 2013, Article ID: 283423.
http://dx.doi.org/10.1155/2013/283423

[16]   Sivaramanan, S. (2014) Isolation of Cellulolytic Fungi and Their Degradation on Cellulosic Agricultural Wastes. Journal of Academia and Industrial Research, 2, 458-463.

[17]   Horner, W.E., Helbling, A., Salvaggio, J.E. and Lehrer, S.B. (1995) Fungal Allergens. Clinical Microbiology Reviews, 8, 161-179.

[18]   Kostylev, M. and Wilson, D. (2012) Synergistic Interactions in Cellulose Hydrolysis. Biofuels, 3, 61-70.
http://dx.doi.org/10.4155/bfs.11.150

[19]   Kim, I.J., Lee, H.J., Choi, I.-G. and Kim, K.H. (2014) Synergistic Proteins for the Enhanced Enzymatic Hydrolysis of Cellulose by Cellulase. Applied Microbiology and Biotechnology, 98, 8469-8480.
http://dx.doi.org/10.1007/s00253-014-6001-3

[20]   Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., Thompson, J.D., Gibson, T.J. and Higgins, D.G. (2007) Clustal W and Clustal X Version 2.0. Bioinformatics, 23, 2947-2948.
http://dx.doi.org/10.1093/bioinformatics/btm404

[21]   Page RDM (1996) TreeView: An Application to Display Phylogenetic Trees on Personal Computers. Computer Applications in the Biosciences, 12, 357-358.

[22]   Hoefer Inc. (1994) Protein Electrophoresis Applications Guide. Hoefer Scientific Instruments, San Francisco.

[23]   Yamanaka, R., Soares, C.F., Matheus, D.R. and Machado, K.M.G. (2008) Lignolytic Enzymes Produced by Trametes villosa CCB176 under Different Culture Conditions. Brazilian Journal of Microbiology, 39, 78-84.
http://dx.doi.org/10.1590/S1517-83822008000100019

[24]   Liu, D.Y., Li, J., Zhao, S., Zhang, R.F., Wang, M.M., Miao, Y.Z., Shen, Y.F. and Shen, Q.R. (2013) Secretome Diversity and Quantitative Analysis of Cellulolytic Aspergillus fumigates Z5 in the Present of Different Carbon Sources. Biotechnology for Biofuels, 6, 149.
http://dx.doi.org/10.1186/1754-6834-6-149

[25]   Ntougias, S., Bourtzis, K. and Tsiamis, G. (2013) The Microbiology of Olive Mill Wastes. BioMed Research International, 2013, Article ID: 784591.
http://dx.doi.org/10.1155/2013/784591

[26]   Nakayan, P., Hameed, A., Singh, S., Young, I.S. and Young, C.C. (2013) Phosphate-Solubilizing Soil Yeast Meyerozyma guilliermondii CC1 Improves Maize (Zea mays L.) Productivity and Minimizes Requisite Chemical Fertilization. Plant and Soil, 373, 301-315.
http://dx.doi.org/10.1007/s11104-013-1792-z

[27]   Kim, J.-S., Baek, J.-H., Park, N.-H. and Kim, C. (2015) Complete Genome Sequence of Halophilic Yeast Meyerozyma caribbica MG20W Isolated from Rhizosphere Soil. Genome Announcements, 3, Article ID: e00127-15.
http://dx.doi.org/10.1128/genomeA.00127-15

[28]   Kim, B.-K., So, J.-H. and Rhee, I.-K. (2011) Molecular Cloning and Functional Expression of an Extracellular Exo-β-(1,3)-Glucanase from Candida fermentati Si. EMBL/GenBank/DDBJ Databases and Universal Protein Resource (UniProt) Databases.
http://www.uniprot.org/uniprot/G5DE31

[29]   Takada, G., Kawaguchi, T., Sumitani, J. and Arai, M. (1998) Cloning, Nucleotide Sequence, and Transcriptional Analysis of Aspergillus aculeatus No. F-50 Cellobiohydrolase I (cbhI) Gene. Journal of Fermentation and Bioengineering, 85, 1-9.
http://dx.doi.org/10.1016/S0922-338X(97)80345-5

[30]   Fujii, T., Fang, X., Inoue, H., Murakami, K. and Sawayama, S. (2009) Enzymatic Hydrolyzing Performance of Acremonium cellulolyticus and Trichoderma reesei against Three Lignocellulosic Materials. Biotechnology for Biofuels, 2, 24.
http://dx.doi.org/10.1186/1754-6834-2-24

[31]   Howard, R.L., Abotsi, E., Jansen van Rensburg, E.L. and Howard, S. (2003) Lignocellulose Biotechnology: Issues of Bioconversion and Enzyme Production. African Journal of Biotechnology, 2, 602-619.
http://dx.doi.org/10.5897/AJB2003.000-1115

[32]   Gao, L., Gao, F., Wang, L.S., Geng, C.L., Chi, L.L., Zhao, J. and Qu, Y.B. (2012) N-Glycoform Diversity of Cellobiohydrolase I from Penicillium decumbens and Synergism of Nonhydrolytic Glycoform in Cellulose Degradation. The Journal of Biological Chemistry, 287, 15906-15915.
http://dx.doi.org/10.1074/jbc.M111.332890

[33]   Mountfort, D.O. and Asher, R.A. (1985) Production and Regulation of Cellulase by Two Strains of the Rumen Anaerobic Fungus Neocallimastix frontalis. Applied and Environmental Microbiology, 49, 1314-1322.

[34]   Boer, H. and Koivula, A. (2003) The Relationship between Thermal Stability and pH Optimum Studied with Wild-Type and Mutant Trichoderma reesei Cellobiohydrolase Cel7A. European Journal of Biochemistry, 207, 841-848.
http://dx.doi.org/10.1046/j.1432-1033.2003.03431.x

[35]   Li, X.L. and Calza, R.E. (1991) Fractionation of Cellulases from the Ruminal Fungus Neocallimastix frontalis EB188. Applied and Environmental Microbiology, 57, 3331-3336.

[36]   Adsul, M.G., Ghule, J.E., Singh, R., Shaikh, H., Bastawde, K.B., Gokhale, D.V. and Varma, A.J. (2004) Polysaccharides from Bagasse: Applications in Cellulase and Xylanase Production. Carbohydrate Polymers, 57, 67-72.
http://dx.doi.org/10.1016/j.carbpol.2004.04.001

[37]   Hao, J.J., Tian, X.J., Song, F.Q., He, X.B., Zhang, Z.J. and Zhang, P. (2006) Involvement of Lignocellulolytic Enzymes in the Decomposition of Leaf Litter in a Subtropical Forest. The Journal of Eukaryotic Microbiology, 53, 193-198.
http://dx.doi.org/10.1111/j.1550-7408.2006.00093.x

[38]   Beldman, G., Searle-van Leeuwen, M.J., Rombouts, F.M. and Voragen, F.G.J. (1985) The Cellulase of Trichoderma viride-Purification, Characterization and Comparison of All Detectable Endoglucanases, Exoglucanases and β-Glucosidases. European Journal of Biochemistry, 146, 301-308.
http://dx.doi.org/10.1111/j.1432-1033.1985.tb08653.x

 
 
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