Secretory expression of β-mannanase in Saccharomyces cerevisiae and its high efficiency for hydrolysis of mannans to mannooligosaccharides.

Applied Microbiology and Biotechnology
Junquan LiuYunhe Cao

Abstract

Degradation of mannans is a key process in the production of foods and prebiotics. β-Mannanase is the key enzyme that hydrolyzes 1,4-β-D-mannosidic linkages in mannans. Heterogeneous expression of β-mannanase in Pichia pastoris systems is widely used; however, Saccharomyces cerevisiae expression systems are more reliable and safer. We optimized β-mannanase gene from Aspergillus sulphureus and expressed it in five S. cerevisiae strains. Haploid and diploid strains, and strains with constitutive promoter TEF1 or inducible promoter GAL1, were tested for enzyme expression in synthetic auxotrophic or complex medium. Highest efficiency expression was observed for haploid strain BY4741 integrated with β-mannanase gene under constitutive promoter TEF1, cultured in complex medium. In fed-batch culture in a fermentor, enzyme activity reached ~ 24 U/mL after 36 h, and production efficiency reached 16 U/mL/day. Optimal enzyme pH was 2.0-7.0, and optimal temperature was 60 °C. In studies of β-mannanase kinetic parameters for two substrates, locust bean gum galactomannan (LBG) gave Km = 24.13 mg/mL and Vmax = 715 U/mg, while konjac glucomannan (KGM) gave Km = 33 mg/mL and Vmax = 625 U/mg. One-hour hydrolysis efficiency values were 57% for 1%...Continue Reading

References

Aug 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·A J BrakeP J Barr
Oct 25, 1981·Journal of Molecular Biology·T P St John, R W Davis
Jan 21, 2000·FEMS Microbiology Reviews·J L Cereghino, J M Cregg
Mar 16, 2002·European Journal of Biochemistry·Bingze XuJan-Christer Janson
Jan 4, 2006·Journal of Biotechnology·Changsoo LeeSeokhwan Hwang
Dec 19, 2007·Critical Reviews in Biotechnology·Samriti Dhawan, Jagdeep Kaur
Apr 4, 2008·Applied Microbiology and Biotechnology·L R S Moreira, E X F Filho
Dec 1, 2004·Nutrition Research Reviews·Glenn R GibsonMarcel B Roberfroid
Feb 9, 2010·Applied Microbiology and Biotechnology·Alimjan IdirisKaoru Takegawa
Mar 3, 2010·Annual Review of Plant Biology·Henrik Vibe Scheller, Peter Ulvskov
Jun 15, 2010·Journal of Biotechnology·Tuan Anh PhamJean-Claude Sigoillot
Jun 30, 2010·Applied Microbiology and Biotechnology·Karen De PourcqNico Callewaert
Sep 14, 2010·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Venkat Reddy PannalaK V Venkatesh
Dec 3, 2010·Applied Microbiology and Biotechnology·Danilo PorroDiethard Mattanovich
Oct 4, 2011·Biotechnology Advances·Eda Celik, Pınar Calık
Dec 2, 2011·FEMS Yeast Research·Nancy A Da Silva, Sneha Srikrishnan
Feb 9, 2012·Applied Microbiology and Biotechnology·Prakram Singh ChauhanNaveen Gupta
Jun 4, 2014·Journal of Industrial Microbiology & Biotechnology·A R MalherbeW H van Zyl
Jul 16, 2014·Critical Reviews in Biotechnology·Montarop YamabhaiDietmar Haltrich
May 15, 2015·Applied Microbiology and Biotechnology·Prakram Singh ChauhanNaveen Gupta
Oct 27, 2015·Bioprocess and Biosystems Engineering·Burcu Gündüz Ergün, Pınar Çalık
Nov 12, 2016·Biotechnology Advances·Praveen Kumar Srivastava, Mukesh Kapoor
Aug 2, 2017·International Journal of Biological Macromolecules·Yan-Xiao LiZheng-Qiang Jiang

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Citations

Jan 2, 2021·Frontiers in Bioengineering and Biotechnology·Aneesa Dawood, Kesen Ma
Jun 3, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yuling MengPiwu Li

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Methods Mentioned

BETA
PCR
glycosylation

Software Mentioned

CBS Prediction Servers
Origin Pro
SnapGene

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