Optimisation of β -Glucosidase Production in a Crude Aspergillus japonicus VIT-SB1 Cellulase Cocktail Using One Variable at a Time and Statistical Methods and its Application in Cellulose Hydrolysis.

Publisher: MDPI Country of Publication: Switzerland NLM ID: 101092791 Publication Model: Electronic Cited Medium: Internet ISSN: 1422-0067 (Electronic) Linking ISSN: 14220067 NLM ISO Abbreviation: Int J Mol Sci Subsets: MEDLINE

Original Publication: Basel, Switzerland : MDPI, [2000-

Cellulose*/metabolism ; Cellulase*/metabolism; beta-Glucosidase/metabolism ; Hydrolysis ; Fermentation ; Glucose

Pulp and paper mill sludge (PPMS) is currently disposed of into landfills which are reaching their maximum capacity. Valorisation of PPMS by enzymatic hydrolysis using cellulases is an alternative strategy. Existing commercial cellulases are expensive and contain low titres of β -glucosidases. In this study, β -glucosidase production was optimised by Aspergillus japonicus VIT-SB1 to obtain higher β -glucosidase titres using the One Variable at a Time (OVAT), Plackett Burman (PBD), and Box Behnken design (BBD)of experiments and the efficiency of the optimised cellulase cocktail to hydrolyse cellulose was tested. β -Glucosidase production was enhanced from 0.4 to 10.13 U/mL, representing a 25.3-fold increase in production levels after optimisation. The optimal BBD production conditions were 6 days of fermentation at 20 °C, 125 rpm, 1.75% soy peptone, and 1.25% wheat bran in (pH 6.0) buffer. The optimal pH for β -glucosidase activity in the crude cellulase cocktail was (pH 5.0) at 50 °C. Optimal cellulose hydrolysis using the crude cellulase cocktail occurred at longer incubation times, and higher substrate loads and enzyme doses. Cellulose hydrolysis with the A. japonicus VIT-SB1 cellulase cocktail and commercial cellulase cocktails resulted in glucose yields of 15.12 and 12.33 µmol/mL glucose, respectively. Supplementation of the commercial cellulase cocktail with 0.25 U/mg of β-glucosidase resulted in a 19.8% increase in glucose yield.

Bioresour Technol. 2018 Nov;267:704-713. (PMID: 30093225)
Nature. 1970 Aug 15;227(5259):680-5. (PMID: 5432063)
Biomolecules. 2019 Jun 06;9(6):. (PMID: 31174354)
Anal Chim Acta. 2009 May 29;642(1-2):171-8. (PMID: 19427473)
J Agric Food Chem. 2011 Mar 9;59(5):1954-61. (PMID: 21294581)
Biomolecules. 2013 Sep 03;3(3):612-31. (PMID: 24970184)
Enzyme Res. 2011;2011:248735. (PMID: 21977312)
Mycology. 2022 Dec 17;14(2):91-107. (PMID: 37152851)
Prep Biochem Biotechnol. 2015 Aug 18;45(6):568-87. (PMID: 25036937)
World J Microbiol Biotechnol. 2014 Jul;30(7):2045-52. (PMID: 24609496)
Mol Biotechnol. 2022 Feb;64(2):130-143. (PMID: 34580813)
3 Biotech. 2016 Dec;6(2):213. (PMID: 28330285)
PeerJ Comput Sci. 2021 Jul 5;7:e623. (PMID: 34307865)
Biotechnol Biofuels. 2016 Mar 31;9:78. (PMID: 27034716)
Biotechnol Rep (Amst). 2017 Jul 22;15:114-124. (PMID: 28794998)
J Hazard Mater. 2020 Dec 5;400:123106. (PMID: 32580093)
Int J Microbiol. 2011;2011:949252. (PMID: 21716658)
Appl Environ Microbiol. 1994 Dec;60(12):4584-6. (PMID: 16349468)
Heliyon. 2020 Dec 07;6(12):e05668. (PMID: 33319112)
J Environ Manage. 2020 Sep 15;270:110914. (PMID: 32721348)
Biotechnol Biofuels. 2019 Nov 02;12:258. (PMID: 31700541)
Foods. 2022 Mar 17;11(6):. (PMID: 35327274)
Sci Rep. 2023 Mar 29;13(1):5134. (PMID: 36991150)
Waste Manag. 2021 Apr 1;124:235-243. (PMID: 33636425)
Food Microbiol. 2021 Dec;100:103859. (PMID: 34416959)
AMB Express. 2020 Oct 8;10(1):182. (PMID: 33030626)
Molecules. 2022 Jan 04;27(1):. (PMID: 35011521)

DST/CON/0177/2018 Technology Innovation Agency

Keywords: Box Behnken design; Plackett Burman design; cellulases; cellulose hydrolysis; endoglucanase; exoglucanase; β-glucosidase

9004-34-6 (Cellulose)
EC 3.2.1.4 (Cellulase)
EC 3.2.1.21 (beta-Glucosidase)
IY9XDZ35W2 (Glucose)

Aspergillus japonicus

Date Created: 20230628 Date Completed: 20230629 Latest Revision: 20230701

20240829

PMC10298073

10.3390/ijms24129928

37373076