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Current Chemical Biology

Editor-in-Chief

ISSN (Print): 2212-7968
ISSN (Online): 1872-3136

Research Article

In-Vitro and In-Silico Characterization of Xylose Reductase from Emericella nidulans

Author(s): Vishal Ahuja*, Aashima Sharma, Ranju Kumari Rathour, Vaishali Sharma, Nidhi Rana and Arvind Kumar Bhatt

Volume 13, Issue 2, 2019

Page: [159 - 170] Pages: 12

DOI: 10.2174/2212796812666180622103906

Price: $65

Abstract

Background: Lignocellulosic residues generated by various anthropogenic activities can be a potential raw material for many commercial products such as biofuels, organic acids and nutraceuticals including xylitol. Xylitol is a low-calorie nutritive sweetener for diabetic patients. Microbial production of xylitol can be helpful in overcoming the drawbacks of traditional chemical production process and lowring cost of production.

Objective: Designing efficient production process needs the characterization of required enzyme/s. Hence current work was focused on in-vitro and in-silico characterization of xylose reductase from Emericella nidulans.

Methods: Xylose reductase from one of the hyper-producer isolates, Emericella nidulans Xlt-11 was used for in-vitro characterization. For in-silico characterization, XR sequence (Accession No: Q5BGA7) was used.

Results: Xylose reductase from various microorganisms has been studied but the quest for better enzymes, their stability at higher temperature and pH still continues. Xylose reductase from Emericella nidulans Xlt-11 was found NADH dependent and utilizes xylose as its sole substrate for xylitol production. In comparison to whole cells, enzyme exhibited higher enzyme activity at lower cofactor concentration and could tolerate higher substrate concentration. Thermal deactivation profile showed that whole cell catalysts were more stable than enzyme at higher temperature. In-silico analysis of XR sequence from Emericella nidulans (Accession No: Q5BGA7) suggested that the structure was dominated by random coiling. Enzyme sequences have conserved active site with net negative charge and PI value in acidic pH range.

Conclusion: Current investigation supported the enzyme’s specific application i.e. bioconversion of xylose to xylitol due to its higher selectivity. In-silico analysis may provide significant structural and physiological information for modifications and improved stability.

Keywords: Xylose reductase, crude enzyme, in silico characterization, Emericella nidulans, lignocellulosic residues, anthropogenic activities.

Graphical Abstract

[1]
Sadh PK, Duhan S, Duhan JS. Agro-industrial wastes and their utilization using solid state fermentation: A review. Bioresour Bioprocess 2018; 5: 1.
[2]
Bhatt AK, Bhalla TC, Agrawal HO, Sharma N. Effect of gamma irradiation on biodegradation of forest lignocelluloses by Aspergillus niger. Biotechnol Tech 1992; 6(2): 111-4.
[3]
Adrio JL, Demain AL. Microbial cells and enzymes a century of progress.In: Barredo JL, Ed.. Methods in biotechnology, microbial enzymes and biotransformations. Totowa, NJ, USA: Humana Press 2015; 17: pp. 1-27.
[4]
Ahuja V, Rathore RK, Bhatia RK, Bhatt AK. Microbial utilization of municipal solid waste (MSW) for the production of Xylitol. Life Sci Inter Res J 2017; 4(1): 56-9.
[5]
Johnson EA. Biotechnology of non-Saccharomyces yeasts-the ascomycetes. Appl Microbiol Biotechnol 2013; 97(2): 503-17.
[6]
Anbu P, Gopinath SCB, Chaulagain BP, Tang TH, Citartan M. Microbial enzymes and their applications in industries and medicine 2014. BioMed Res Int 2015; 816419: 3.
[7]
Kumar S, Dhankhar S, Arya VP, Yadav S, Yadav JP. Antimicrobial activity of Salvadora oleoides Decne. against some microorganisms. J Med Plants Res 2013; 6(14): 2754-60.
[8]
Liu X, Kokare C. Microbial enzymes of use in industry.In: Brahmachari G. Eds.. Biotechnology of microbial enzymes. 2016; pp. 267-98.
[9]
de Carvalho CCCR. Whole cell biocatalysts: Essential workers from nature to the industry. Microb Biotechnol 2016; 10(2): 250-63.
[10]
Mussatto SI, Roberto IC. Alternatives for detoxification of diluted acid lignocellulosic hydrolysates for use in fermentative processes: A review. Bioresour Technol 2014; 93(1): 1-10.
[11]
de Sousa LP, da Silva AF, Calil NO, Oliveira MG, da Silva SS. Raposo NRB. In-vitro inhibition of Pseudomonas aeruginosa adhesion by xylitol. Braz Arch Biol Technol 2011; 54(5): 877-84.
[12]
Rehman S, Mushtaqm Z, Zahoor T, Jamil A, Murtaza MA. Xylitol: A review on bioproduction, application, health benefits, and related safety issues. Crit Rev Food Sci Nutr 2015; 55(11): 1514-28.
[13]
Amo K, Arai H, Uebanso T, et al. Effects of xylitol on metabolic parameters and visceral fat accumulation. J Clin Biochem Nutr 2011; 49(1): 1-7.
[14]
Verduyn C, Van Kleef R, Frank J, Schreuder H, Van Dijken JP, Scheffers WA. Properties of the NAD(P)H-dependent xylose reductase from the xylose-fermenting yeast Pichia stipites. Biochem J 1985; 226(3): 669-77.
[15]
Woodyer R, Simurdiak M, Van Der Donk WA, Zhao H. Heterogeneous expression, purification, and characterization of a highly active xylose reductase from Neurospora crassa. Appl Environ Microbiol 2005; 71(3): 1642-7.
[16]
Rafiqul ISM, Sakinah M. Bioproduction of xylitol by enzyme technology and future prospects. Int Food Res J 2012; 19(2): 405-8.
[17]
Yokoyama SI, Suzuki T, Kawai K, Horitsu H, Takamizawa K. Purification, characterization and structure analysis of NADPH-Dependent D-Xylose reductase from Candida tropicalis. J Ferment Bioeng 1995; 79(3): 217-23.
[18]
Geourjon C, Deleage G. SOPM: A self-optimized method for protein secondary structure prediction. Protein Eng 1994; 7(2): 157-64.
[19]
Ma J, Wang S, Zhao F, Xu J. Protein threading using context-specific alignment potential. Bioinformatics 2013; 29(13): i257-65.
[20]
Guo J, Huang S, Chen Y, Guo X, Xiao D. Heterologous expression of Spathaspora Passalidarum xylose reductase and xylitol dehydrogenase genes improved xylose fermentation ability of Aureobasidium pullulans. Microb Cell Fact 2018; 17(1): 64.
[21]
Sathesh-Prabu C, Lee SK. Enhancement of α,ω-dicarboxylic acid production by the expression of xylose reductase for refactoring redox cofactor regeneration. J Agric Food Chem 2018; 66(13): 3489-97.
[22]
Su Y, Li W, Zhu W, et al. Characterization of xylose reductase from Candida tropicalis immobilized on chitosan bead. Afr J Biotechnol 2010; 9(31): 4954-65.
[23]
Malla S, Gummadi SN. Thermal stability of xylose reductase from Debaryomyces nepalensis NCYC 3413: Deactivation kinetics and structural studies. Process Biochem 2018; 67: 71-9.
[24]
Komeda H, Yamasaki-Yashiki S, Hoshino K, Asano Y. Identification and characterization of d-xylose reductase involved in pentose catabolism of the zygomycetous fungus Rhizomucor pusillus. J Biosci Bioeng 2015; 119(1): 57-64.

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