Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

doi:10.1016/j.cjche.2019.11.012

Chinese Journal of Chemical Engineering ›› 2020, Vol. 28 ›› Issue (4): 1120-1128.DOI: 10.1016/j.cjche.2019.11.012

• Biotechnology and Bioengineering • Previous Articles Next Articles

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

Linian Cai¹, Shengnan Xu¹, Tao Lu², Dongqiang Lin¹, Shanjing Yao¹

1 Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2 College of Environment, Zhejiang University of Technology, Hangzhou 310014, China

Received:2019-10-10 Revised:2019-11-21 Online:2020-07-27 Published:2020-04-28
Contact: Linian Cai, Shengnan Xu, Tao Lu, Dongqiang Lin, Shanjing Yao
Supported by:
This work was supported by National Natural Science Foundation of China (21576233, 21878263) and Fundamental Research Funds for the Central Universities.

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

Linian Cai¹, Shengnan Xu¹, Tao Lu², Dongqiang Lin¹, Shanjing Yao¹

1 Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2 College of Environment, Zhejiang University of Technology, Hangzhou 310014, China

通讯作者: Linian Cai, Shengnan Xu, Tao Lu, Dongqiang Lin, Shanjing Yao
基金资助:
This work was supported by National Natural Science Foundation of China (21576233, 21878263) and Fundamental Research Funds for the Central Universities.

Abstract

Abstract: The cellulase cocktail produced by marine Aspergillus niger exhibits a property of salt-tolerance, which is of great potential in cellulose degradation in high salt environment. In order to explain the mechanism on the salttolerance of the cellulase cocktail produced by marine A. niger, six cellulase components (AnCel6, AnCel7A, AnCel7B, AnEGL, AnBGL1 and AnBGL2) were obtained by directed expression. Studies on their enzymatic properties revealed that one β-glucosidase (AnBGL2) and one endoglucanase (AnEGL) exhibited an outstanding salttolerant property, and one cellobiohydrolase (AnCel7B) exhibited a certain salt-tolerant property. Subsequent study revealed that the salt-tolerant AnEGL and AnCel7B endowed the cellulase cocktail with stronger salttolerant property, while the salt-tolerant AnBGL2 had no positive effect. Moreover, after overexpression of AnCel6, AnCel7A, AnCel7B and AnEGL, the activity of cellulase cocktail increased by 80%, 70%, 63% and 68%, respectively. However, the activity of cellulase cocktail was not improved after overexpression of AnBGL1 and AnBGL2. After mixed-strain fermentation with cellobiohydrolase recombinants (cel6a, cel7a and cel7b recombinants) and endoglucanase recombinant (egl recombinant), the the activity of cellulase cocktail increased by 114%, 102% and 91%, respectively.

Key words: Marine Aspergillus niger, Cellulase component, Directed expression, Salt-tolerance, Cellulase activity

摘要： The cellulase cocktail produced by marine Aspergillus niger exhibits a property of salt-tolerance, which is of great potential in cellulose degradation in high salt environment. In order to explain the mechanism on the salttolerance of the cellulase cocktail produced by marine A. niger, six cellulase components (AnCel6, AnCel7A, AnCel7B, AnEGL, AnBGL1 and AnBGL2) were obtained by directed expression. Studies on their enzymatic properties revealed that one β-glucosidase (AnBGL2) and one endoglucanase (AnEGL) exhibited an outstanding salttolerant property, and one cellobiohydrolase (AnCel7B) exhibited a certain salt-tolerant property. Subsequent study revealed that the salt-tolerant AnEGL and AnCel7B endowed the cellulase cocktail with stronger salttolerant property, while the salt-tolerant AnBGL2 had no positive effect. Moreover, after overexpression of AnCel6, AnCel7A, AnCel7B and AnEGL, the activity of cellulase cocktail increased by 80%, 70%, 63% and 68%, respectively. However, the activity of cellulase cocktail was not improved after overexpression of AnBGL1 and AnBGL2. After mixed-strain fermentation with cellobiohydrolase recombinants (cel6a, cel7a and cel7b recombinants) and endoglucanase recombinant (egl recombinant), the the activity of cellulase cocktail increased by 114%, 102% and 91%, respectively.

关键词: Marine Aspergillus niger, Cellulase component, Directed expression, Salt-tolerance, Cellulase activity

Linian Cai, Shengnan Xu, Tao Lu, Dongqiang Lin, Shanjing Yao. Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity[J]. Chinese Journal of Chemical Engineering, 2020, 28(4): 1120-1128.

Linian Cai, Shengnan Xu, Tao Lu, Dongqiang Lin, Shanjing Yao. Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity[J]. 中国化学工程学报, 2020, 28(4): 1120-1128.

References

[1] C.M. Payne, B.C. Knott, H.B. Mayes, H. Hansson, M.E. Himmel, M. Sandgren, J. Ståhlberg, G.T. Beckham, Fungal cellulases, Chem. Rev. 115(2015) 1308-1448.
[2] S.M. Cragg, G.T. Beckham, N.C. Bruce, T.D. Bugg, D.L. Distel, P. Dupree, A.G. Etxabe, B. S. Goodell, J. Jellison, J.E. McGeehan, S.J. McQueen-Mason, K. Schnorr, P.H. Walton, J. E. Watts, M. Zimmer, Lignocellulose degradation mechanisms across the tree of life, Curr. Opin. Chem. Biol. 29(2015) 108-119.
[3] L. Du, L. Ma, Q. Ma, G. Guo, X. Han, D. Xiao, Hydrolytic boosting of lignocellulosic biomass by a fungal lytic polysaccharide monooxygenase, AnLPMO15g from Aspergillus niger, Ind. Crop. Prod. 126(2018) 309-315.
[4] S.K. Soni, A. Sharma, R. Soni, Cellulases:Role in lignocellulosic biomass utilization, in:M. Lübeck (Ed.), Cellulases, Humana Press, New Jersey, 2018.
[5] Z. Zhou, F. Lei, P. Li, J. Jiang, Lignocellulosic biomass to biofuels and biochemicals:A comprehensive review with a focus on ethanol organosolv pretreatment technology, Biotechnol. Bioeng. 115(2018) 2683-2702.
[6] H.B. Lei, J. Wang, Study on the mechanism of improving the digestibility of ruminant to roughage fiber by different processing methods, Journal of Animal Science and Veterinary Medicine 38(2019) 50-52.
[7] R. Liu, Q. Han, Q.Q. Qian, X.W. Peng, Process optimization and analysis of hydrolyzate in paper mill sludge hydrolysis with cellulase, China Pulp & Paper 35(2016) 42-46.
[8] J.Xue,X.G.Li,J.Z.Wang,F.J.Wang,Optimizationofenzymatichydrolysisforproduction Aronia melanocarpa juice and functional properties, Sci. Technol. Food Ind. (2019) 1-17.
[9] Y.Y. Yu, Q. Wang, X.R. Fan, Application of cellulase for cellulosic fabrics finishing, Textile Dyeing and Finishing Journal 39(2017) 1-5.
[10] R. Peterson, H. Nevalainen, Trichoderma reesei RUT-C30-thirty years of strain improvement, Microbiology 158(2011) 58-68.
[11] D.S. Xue, H.Y. Chen, D.Q. Lin, Y.X. Guan, S.J. Yao, Optimization of a natural medium for cellulase by a marine Aspergillus niger using response surface methodology, Appl. Biochem. Biotechnol. 167(2012) 1963-1972.
[12] D.U. Jung, H.Y. Yoo, S.B. Kim, J.H. Lee, C. Park, S.W. Kim, Optimization of medium composition for enhanced cellulase production by mutant Penicillium brasilianum KUEB15 using statistical method, J. Ind. Eng. Chem. 25(2015) 145-150.
[13] A. Sørensen, M. Lübeck, P. Lübeck, B. Ahring, Fungal beta-glucosidases:A bottleneck in industrial use of lignocellulosic materials, Biomolecules 3(2013) 612-631.
[14] R.R. Singhania, R.K. Sukumaran, A.K. Patel, C. Larroche, A. Pandey, Advancement and comparative profiles in the production technologies using solid-state and submerged fermentation for microbial cellulases, Enzym. Microb. Technol. 46(2010) 541-549.
[15] J.J. Cai, X.H. Duan, L. Xie, X.F. Zheng, J.T. Shen, Production of cellulases by solid state fermentation with three strains mixed, China Biotechnology 33(2013) 57-63.
[16] X. Tu, Q.H. Xue, M.R. Si, M.F. Gong, Effects of mixed poly-fermentation on cellulase activity, Industrial Microbiology (2004) 30-34.
[17] B.B. Wang, L.M. Xia, High efficient expression of cellobiase gene from Aspergillus niger in the cells of Trichoderma reesei, Bioresour. Technol. 102(2011) 4568-4572.
[18] M. Dashtban, W. Qin, Overexpression of an exotic thermotolerant beta-glucosidase in Trichoderma reesei and its significant increase in cellulolytic activity and saccharification of barley straw, Microb. Cell Factories 11(2012) 63.
[19] H. Fang, L. Xia, High activity cellulase production by recombinant Trichoderma reesei ZU-02 with the enhanced cellobiohydrolase production, Bioresour. Technol. 144(2013) 693-697.
[20] D.S. Xue, L.Y. Liang, G. Zheng, D.Q. Lin, Q.L. Zhang, S.J. Yao, Expression of Piromyces rhizinflata cellulase in marine Aspergillus niger to enhance halostable cellulase activity by adjusting enzyme-composition, Biochem. Eng. J. 117(2017) 156-161.
[21] C.P. Kubicek, M. Mikus, A. Schuster, M. Schmoll, B. Seiboth, Metabolic engineering strategies for the improvement of cellulase production by Hypocrea jecorina, Biotechnol. Biofuels 2(2009) 19.
[22] T. Nakari-Setala, M. Paloheimo, J. Kallio, J. Vehmaanpera, M. Penttila, M. Saloheimo, Genetic modification of carbon catabolite repression in Trichoderma reesei for improved protein production, Appl. Environ. Microbiol. 75(2009) 4853-4860.
[23] N. Aro, M. Ilmen, A. Saloheimo, M. Penttila, ACEI of Trichoderma reesei is a repressor of cellulase and xylanase expression, Appl. Environ. Microbiol. 69(2003) 56-65.
[24] S. DasSarma, P. DasSarma, Halophiles and their enzymes:Negativity put to good use, Curr. Opin. Microbiol. 25(2015) 120-126.
[25] X.Q. Wang, X. Xing, F.J. Zhang, K. Xin, Biological improvement of saline alkali soil reference system:A review, Sciences in Cold and Arid Regions 10(2018) 516-521.
[26] M. Galbe, G. Zacchi, Pretreatment of lignocellulosic materials for efficient bioethanol production, in:L. Olsson (Ed.), Biofuels, Springer, Berlin, 2007.
[27] A. Brandt, M.J. Ray, T.Q. To, D.J. Leak, R.J. Murphy, T. Welton, Ionic liquid pretreatment of lignocellulosic biomass with ionic liquid-water mixtures, Green Chem. 13(2011) 2489.
[28] D.S. Xue, Production and Proerties of Salt-Tolerant Cellulase and β-Glucosidase from a Marine Aspergillus niger, Ph. D. Thesis, Zhejiang Univ., China, 2012.
[29] D.M. Bai, Removal and Mechanism of Ni(Ⅱ) and Cr(VI) from Aqueous Solutions by Marine Aspergillus niger Mycelial Pellet, M. Phil. Thesis, Zhejiang Univ., China, 2016.
[30] T. Lu, The Pellet-Formation Mechanism of Marine-Derived Aspergillus niger and its Application inDye-WastewaterTreatment, Ph.D.Thesis, ZhejiangUniv.,China,2016.
[31] C.B. Michielse, P.J. Hooykaas, C.A. van den Hondel, A.F. Ram, Agrobacterium-mediated transformation of the filamentous fungus Aspergillus awamori, Nat. Protoc. 3(2008) 1671-1678.
[32] A.A. Wise, Z. Liu, A.N. Binns, Three methods for the introduction of foreign DNA into Agrobacterium, Methods Mol. Biol. 343(2006) 43-53.
[33] N.J. Kruger, The Bradford method for protein quantitation, the protein protocols handbook, in:J.M. Walker (Ed.), The Protein Protocols Handbook, Humana Press, New Jersey, 2009.
[34] T.M. Wood, K.M. Bhat, Methods for measuring cellulase activity, Methods Enzymol. 160(1988) 87-112.
[35] L.N. Cai, S.N. Xu, T. Lu, D.Q. Lin, S.J. Yao, Directed expression of halophilic and acidophilic β-glucosidases by introducing homologous constitutive expression cassettes in marine Aspergillus niger, J. Biotechnol. 292(2019) 12-22.
[36] R. Baghban, S. Farajnia, M. Rajabibazl, Y. Ghasemi, A. Mafi, R. Hoseinpoor, L. Rahbarnia, M. Aria, Yeast expression systems:Overview and recent advances, Mol. Biotechnol. 61(2019) 365-384.
[37] A. Tsang, G. Butler, J. Powlowski, E.A. Panisko, S.E. Baker, Analytical and computational approaches to define the Aspergillus niger secretome, Fungal Genet. Biol. 46(2009) S153-S160.
[38] L. Ma, J. Zhang, G. Zou, C. Wang, Z. Zhou, Improvement of cellulase activity in Trichoderma reesei by heterologous expression of a beta-glucosidase gene from Penicillium decumbens, Enzym. Microb. Technol. 49(2011) 366-371.
[39] H. Nakazawa, T. Kawai, N. Ida, Y. Shida, Y. Kobayashi, H. Okada, S. Tani, J. Sumitani, T. Kawaguchi, Y. Morikawa, W. Ogasawara, Construction of a recombinant Trichoderma reesei strain expressing Aspergillus aculeatus beta-glucosidase 1 for efficient biomass conversion, Biotechnol. Bioeng. 109(2012) 92-99.
[40] G.B. Sperandio, E.X. Ferreira Filho, Fungal co-cultures in the lignocellulosic biorefinery context:A review, Int. Biodeterior. Biodegradation 142(2019) 109-123.

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

Salt-tolerant mechanism of marine Aspergillus niger cellulase cocktail and improvement of its activity

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