Advances in the study of directed evolution for cellulases

doi:10.1007/s11783-011-0326-2

Front Envir Sci Eng Chin

2011, Vol. 5

Issue (4) : 519-525 https://doi.org/10.1007/s11783-011-0326-2

RESEARCH ARTICLE

Advances in the study of directed evolution for cellulases

Hailong LIN^1,2, Weiguang LI¹, Changhong GUO², Sihang QU², Nanqi REN¹(

)

1. State Key Laboratory of Urban Water Resource and Environment, Harbin Institute of Technology, Harbin 150090, China; 2. College of Life Science and Technology, Harbin Normal University, Harbin 150080, China

Download: PDF(117 KB) HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks

Abstract

If cellulose can be effectively hydrolyzed into glucose by cellulase, the production costs of hydrogen, ethanol or other chemicals from cellulosic materials will be greatly decreased, and economically viable production of biohydrogen and bioethanol will become feasible. Cellulose is degraded into glucoses by multi-component enzyme systems. Nowadays cellulases are widely used in brewing, food, bioenergy, fodder, textiles, paper, pharmaceuticals, environmental protection and other industries. However, existing cellulases have several problems that limit their wider applications, including the low turnover number for solid cellulosic materials, and low stability in adapting to various application conditions. For example, high temperature, low pH, and so on. Application of directed evolution technology may be one of the most effective ways for improving the characteristics of cellulases. This paper presents a brief review of the cellulose hydrolysis mechanism by cellulase, advances in cellulases (endoglucanase and β-glucosidase) improvement by directed evolution for several characteristics (for instance, thermal stability, pH adaptability and enzyme activity), limitations of directed evolution for cellulases, and the outlook for directed evolution for cellulase.

Keywords biohydrogen bioethanol cellulase cellulose directed evolution

Corresponding Author(s): REN Nanqi,Email:rnq@hit.edu.cn

Issue Date: 05 December 2011

Cite this article:

Hailong LIN,Weiguang LI,Changhong GUO, et al. Advances in the study of directed evolution for cellulases[J]. Front Envir Sci Eng Chin, 2011, 5(4): 519-525.

URL:

https://academic.hep.com.cn/fese/EN/10.1007/s11783-011-0326-2
https://academic.hep.com.cn/fese/EN/Y2011/V5/I4/519

enzyme	altered property	DNA technique	screening/selection	expression system	Ref.
endoglucanase	thermal stability	family shuffling	facilitated screening-Congo red + CMC agar	E. coli	[15,32]
endoglucanase	activity	DNA shuffling	facilitated screening-Congo red + CMC agar	E. coli (cell surface display)	[16,32]
endoglucanase	none	family shuffling	facilitated screening-Congo red + CMC agar	Neurospora spheroplasts	[17,32]
β-D-glucosidase	cold adaption	DNA shuffling	random screening-chromogenic substrate	yeast	[18,32]
β-glycosidase	activity	family shuffling	random screening-chromogenic substrate	E. coli	[19,32]
endoglucanase	alkali pH	epPCR	facilitated screening-Congo red + CMC agar	yeast	[20,32]
β-D -glucosidase	thermal stability	epPCR	random screening-chromogenic substrate	E. coli	[21,33]
β-D -glucosidase	activity	epPCR	random screening-coupled to color reaction	E. coli	[22,32]
mutated β-glucosidase(glycosynthase)	activity	epPCR	facilitated screening-fluorogenic substrate	E. coli	[23,32]
endoglucanase	stability, activity	epPCR	facilitated screening-Congo red + CMC agar	E. coli	[24]
endoglucanase	cold adaptation	epPCR	facilitated screening-Congo red + CMC agar	yeast	[25]
endoglucanase	catalytic efficiency and pH	epPCR and DNA shuffling	facilitated screening-Congo red + CMC agar	yeast	[26]
β-D-glucosidase	thermal stability	epPCR + familyshuffling	random screening-chromogenic substrate	E.coli	[27,32]
endo-β-1, 4-glucanase	activity and thermal stability and pH	DNA shuffling	facilitated screening-Congo red + CMC agar	E.coli	[28]
endoglucanase	thermal stability pH	epPCR + familyshuffling	facilitated screening-Congo red + CMC agar	E.coli	[29]
endoglucanase	activity, pH, stability	epPCR + family shuffling	facilitated screening-Congo red + CMC agar	yeast	[30]
β-D-glucosidase	thermal stability	epPCR	M9 + Cellobiose + indicator strain	E.coli	[33]
cellobiohydrolase	thermal stability	SCHEMA	monomera screening strategy	yeast	[37]
cellobiohydrolase	thermal stability	SCHEMA	monomera screening strategy	yeast	[38]
β-D -glucosidase	catalytic efficiency	epPCR	fluorescence-activated cell sorting (FACS)	E.coli	[39]

Tab.1 Cellulases and relevant enzymes whose properties have been changed using directed evolution techniques

1	Hammerschlag R. Ethanol’s energy return on the investment: a survey of the literature 1990–present. Environmental Science & Technology , 2006, 40(6): 1744–1750 doi: 10.1021/es052024h
2	Lynd L R, Laser M S, Bransby D, Dale B E, Davison B, Hamilton R, Himmel M, Keller M, McMillan J D, Sheehan J, Wyman C E. How biotech can transform biofuels. Nature Biotechnology , 2008, 26(2): 169–172 doi: 10.1038/nbt0208-169
3	Alriksson B, Rose S H, van Zyl W H, Sjode A, Nilvebrant N O, Jonsson L J. Cellulase production from spent lignocellulose hydrolysates by recombinant Aspergillus niger. Applied and Environmental Microbiology , 2009, 75(8): 2366–2374 doi: 10.1128/AEM.02479-08
4	Vrije T, Bakker R R, Budde M A W, Lai M H, Mars A E, Claassen P A M. Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana. Biotechnology for Biofuels , 2009, 2(1): 12–26 doi: 10.1186/1754-6834-2-12
5	Dashtban M, Schraft H, Qin W S. Fungal bioconversion of lignocellulosic residues: opportunities & perspectives. International Journal of Biological Sciences , 2009, 5(6): 578–595
6	Maki M, Leung K T, Qin W S. The prospects of cellulase-producing bacteria for the bioconversion of lignocellulosic biomass. International Journal of Biological Sciences , 2009, 5(5): 500–516
7	Hu L Y, Zhong W H. Advances of the research on gene cloning and functional amino-acid of cellulose. Biotechnology China , 2003, 13(2): 43–45 (in Chinese)
8	Taherzadeh M J, Karimi K. Pretreatment of lignocellulosic wastes to improve ethanol and biogas production: a review. International Journal of Molecular Sciences , 2008, 9(9): 1621–1651 doi: 10.3390/ijms9091621
9	Lynd L R, Weimer P J, van Zyl W H, Pretorius I S. Microbial cellulose utilization: fundamentals and biotechnology. Microbiology and Molecular Biology Reviews , 2002, 66(3): 506–577 doi: 10.1128/MMBR.66.3.506-577.2002
10	Demain A L, Newcomb M, Wu J H D. Cellulase, clostridia, and ethanol. Microbiology and Molecular Biology Reviews , 2005, 69(1): 124–154 doi: 10.1128/MMBR.69.1.124-154.2005
11	Yu X L, Wang L, Xu W M. Progress in the studies of cellulose degradation by cellulase. Journal of Ningbo University , 2007, 20(1): 78–82 (in Chinese)
12	Eijsink V G H, G?seidnes S, Borchert T V, van den Burg B. Directed evolution of enzyme stability. Biomolecular Engineering , 2005, 22(1–3): 21–30 doi: 10.1016/j.bioeng.2004.12.003
13	Hibbert E G, Dalby P A. Directed evolution strategies for improved enzymatic performance. Microbial Cell Factories , 2005, 4(1): 29–34 doi: 10.1186/1475-2859-4-29
14	Liu Y, Zhang H F, Sun Z. Advance in molecular biology and gene engineer of cellulase. Feed industry China , 2007, 28 (18): 11–14 (in Chinese)
15	Murashima K, Kosugi A, Doi R H. Thermostabilization of cellulosomal endoglucanase EngB from Clostridium cellulovorans by in vitro DNA recombination with non-cellulosomal endoglucanase EngD. Molecular Microbiology , 2002, 45(3): 617–626 doi: 10.1046/j.1365-2958.2002.03049.x
16	Kim Y S, Jung H C, Pan J G. Bacterial cell surface display of an enzyme library for selective screening of improved cellulase variants. Applied and Environmental Microbiology , 2000, 66(2): 788–793 doi: 10.1128/AEM.66.2.788-793.2000
17	Catcheside D E A, Rasmussen J P, Yeadon P J, Bowing F J, Cambareri E B, Kato E, Gabe J, Stuart W D. Diversification of exogenous genes in vivo in Neurospora. Applied and Environmental Microbiology , 2003, 62: 544–549
18	Lebbink J H, Kaper T, Bron P, van der Oost J, de Vos W M. Improving low-temperature catalysis in the hyperthermostable Pyrococcus furiosus beta-glucosidase CelB by directed evolution. Biochemistry , 2000, 39(13): 3656–3665 doi: 10.1021/bi991483q
19	Kaper T, Brouns S J J, Geerling A C M, de Vos W M, van der OOST J. DNA family shuffling of hyperthermostable beta-glycosidases. Biochemical Journal , 2002, 368(2): 461–470 doi: 10.1042/BJ20020726
20	Wang T, Liu X, Yu Q, Zhang X, Qu Y, Gao P, Wang T. Directed evolution for engineering pH profile of endoglucanase III from Trichoderma reesei. Biomolecular Engineering , 2005, 22(1–3): 89–94 doi: 10.1016/j.bioeng.2004.10.003
21	González-Blasco G, Sanz-Aparicio J, González B, Hermoso J A, Polaina J. Directed evolution of beta-glucosidase A from Paenibacillus polymyxa to thermal resistance. Journal of Biological Chemistry , 2000, 275(18): 13708–13712 doi: 10.1074/jbc.275.18.13708
22	McCarthy J K, Uzelac A, Davis D F, Eveleigh D E. Improved catalytic efficiency and active site modification of 1,4-beta-D-glucan glucohydrolase A from Thermotoga neapolitana by directed evolution. Journal of Biological Chemistry , 2004, 279(12): 11495–11502 doi: 10.1074/jbc.M305642200
23	Kim Y W, Lee S S, Warren R A, Withers S G. Directed evolution of a glycosynthase from Agrobacterium sp. increases its catalytic activity dramatically and expands its substrate repertoire. Journal of Biological Chemistry , 2004, 279(41): 42787–42793 doi: 10.1074/jbc.M406890200
24	Nakazawa H, Okada K, Onodera T, Ogasawara W, Okada H, Morikawa Y. Directed evolution of endoglucanase III (Cel12A) from Trichoderma reesei. Applied Microbiology and Biotechnology , 2009, 83(4): 649–657 doi: 10.1007/s00253-009-1901-3
25	Xiao Z Z, Wang P, Qu Y B, Gao P J, Wang T H. Cold adaptation of a mesophilic cellulase, EG III from Trichoderma reesei, by directed evolution. Science in China. Series C, Life Sciences , 2002, 45(4): 337–343 doi: 10.1360/02yc9037
26	Qin Y Q, Wei X M, Song X, Qu Y B. Engineering endoglucanase II from Trichoderma reesei to improve the catalytic efficiency at a higher pH optimum. Journal of Biotechnology , 2009, 135(2): 190–195 doi: 10.1016/j.jbiotec.2008.03.016
27	Arrizubieta M J, Polaina J. Increased thermal resistance and modification of the catalytic properties of a beta-glucosidase by random mutagenesis and in vitro recombination. Journal of Biological Chemistry , 2000, 275(37): 28843–28848 doi: 10.1074/jbc.M003036200
28	Lin L, Meng X, Liu P F, Hong Y Z, Wu G B, Huang X L, Li C C, Dong J L, Xiao L, Liu Z. Improved catalytic efficiency of Endo-β-1, 4-glucanase from Bacillus subtilis BME-15 by directed evolution. Applied Microbiology and Biotechnology , 2009, 82(4): 671–679 doi: 10.1007/s00253-008-1789-3
29	Heinzelman P, Komor R, Kanaan A, Romero P, Yu X L, Mohler S, Snow C, Arnold F. Efficient screening of fungal cellobiohydrolase classIenzymes for thermostabilizing sequence blocks by SCHEMA structure-guided recombination. Protein Engineering, Design & Selection , 2010, 23(11): 871–880 doi: 10.1093/protein/gzq063
30	Heinzelman P, Snow C D, Smith M A, Yu X L, Kannan A, Boulware K, Villalobos A, Govindarajan S, Minshull J, Arnold F H. SCHEMA recombination of a fungal cellulase uncovers a single mutation that contributes markedly to stability. Journal of Biological Chemistry , 2009, 284(39): 26229–26233 doi: 10.1074/jbc.C109.034058
31	Zhang X Y. Studies on the directed evolution and thermostability of β-glucanase. Dissertation for Doctoral Degree . Hangzhou: Zhejiang University, 2006, 64–94 (in Chinese)
32	Qin Y Q. Protein engineering and expression of the endoglucanase II from Trichoderma reesei. Dissertation for Doctoral Degree . Jinan: Shandong University, 2008: 71–101 (in Chinese)
33	Hardiman E, Gibbs M, Reeves R, Bergquist P. Directed evolution of a thermophilic β-glucosidase for cellulosic bioethanol production. Applied Biochemistry Biotechnology , 2010 161(1–8):301–312
34	Zhao L G, Meng P, Li L J, Yu S Y. Detection and identification of β-glucosidase with esculin as substrste. Food and Fermentation Industries , 2008, 34(12): 163–166 (in Chinese)
35	Liu W, Hong J, Bevan D R, Zhang Y H P. Fast identification of thermostable beta-glucosidase mutants on cellobiose by a novel combinatorial selection/screening approach. Biotechnology and Bioengineering , 2009, 103(6): 1087–1094 doi: 10.1002/bit.22340
36	Dashtban M, Maki M, Leung K T, Mao C Q, Qin W S. Cellulase activities in biomass conversion: measurement methods and comparison. Critical Reviews in Biotechnology, 2010, 30 (4): 302–309
37	Sen S, Dasu D V, Mandal B. Developments in directed evolution for improving enzyme functions. Applied Biochemistry and Biotechnology , 2007, 143(3): 212–223 doi: 10.1007/s12010-007-8003-4
38	Percivalzhang Y, Himmel M, Mielenz J. Outlook for cellulase improvement: screening and selection strategies. Biotechnology Advances , 2006, 24(5): 452–481 doi: 10.1016/j.biotechadv.2006.03.003
39	Nakazawa H, Okada K, Kobayashi R, Kubota T, Onodera T, Ochiai N, Omata N, Ogasawara W, Okada H, Morikawa Y. Characterization of the catalytic domains of Trichoderma reesei endoglucanase I, II and III, expressed inEscherichia coli. Applied Microbiology and Biotechnology , 2008, 81(4): 681–689 doi: 10.1007/s00253-008-1667-z

[1]	Haifa RAJHI,Daniel PUYOL,Mirna C. MARTÍNEZ,Emiliano E. DÍAZ,José L. SANZ. Vacuum promotes metabolic shifts and increases biogenic hydrogen production in dark fermentation systems[J]. Front. Environ. Sci. Eng., 2016, 10(3): 513-521.
[2]	Ruiqiang LIU,Rattan LAL. Effects of molecular weight and concentration of carboxymethyl cellulose on morphology of hydroxyapatite nanoparticles as prepared with one-step wet chemical method[J]. Front. Environ. Sci. Eng., 2015, 9(5): 804-812.
[3]	Guochen ZHENG, Jianzheng LI, Feng ZHAO, Liguo ZHANG, Li WEI, Qiaoying BAN, Yongsheng ZHAO. Effect of illumination on the hydrogen-production capability of anaerobic activated sludge[J]. Front Envir Sci Eng, 2012, 6(1): 125-130.
[4]	Guanghong ZHENG, Zhuhui KANG, Yifan QIAN, Lei WANG, . Enhanced biohydrogen generation from organic wastewater containing N H 4 + by phototrophic bacteria Rhodobacter sphaeroides AR-3[J]. Front.Environ.Sci.Eng., 2009, 3(4): 387-392.
[5]	Bo WANG, Wei WAN, Jianlong WANG, . Effects of nitrate concentration on biological hydrogen production by mixed cultures[J]. Front.Environ.Sci.Eng., 2009, 3(4): 380-386.

Viewed

Full text

Abstract

Cited

Shared

Discussed