High Production of Trametes cinnabarina Laccase (lac1) by Suspended and Immobilized Cells of Recombinant Pichia pastoris from Crude Glycerol

Bhattacharyya, A; Ahmed, M; Wadhwa, R; Aggarwal, S; Mustafiz, A; Rajagopalan, G

High Production of Trametes cinnabarina Laccase (lac1) by Suspended and Immobilized Cells of Recombinant Pichia pastoris from Crude Glycerol

Bhattacharyya, A Ahmed, M Wadhwa, R Aggarwal, S Mustafiz, A Rajagopalan, G

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Publisher:: Springer
Publication Type:: Journal Article
Citation:: Waste and Biomass Valorization, 2022, 13, (4), pp. 2149-2168
Issue Date:: 2022-04-01

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Field	Value	Language
dc.contributor.author	Bhattacharyya, A
dc.contributor.author	Ahmed, M
dc.contributor.author	Wadhwa, R
dc.contributor.author	Aggarwal, S
dc.contributor.author	Mustafiz, A
dc.contributor.author	Rajagopalan, G
dc.date.accessioned	2022-11-09T04:05:15Z
dc.date.available	2022-11-09T04:05:15Z
dc.date.issued	2022-04-01
dc.identifier.citation	Waste and Biomass Valorization, 2022, 13, (4), pp. 2149-2168
dc.identifier.issn	1877-2641
dc.identifier.issn	1877-265X
dc.identifier.uri	http://hdl.handle.net/10453/163367
dc.description.abstract	Laccases are green catalysts widely employed in various industrial, environmental, clinical and biotechnological applications. This study was aimed to develop the cost effective and efficient process for high level of fungal laccase production from crude glycerol. Initially, laccase gene (lac1) of Trametes cinnabarina was chemically synthesized, and integrated into Pichia pastoris strains (BG10, 11 and 16) by using pD915 vector which can facilitate the constitutive secretory expression of laccase (lac1). Among the recombinant P. pastoris strains developed, st.BG10 (wild type) was supported the high level of laccase production (2851 ± 72 U/l) from pure glycerol based media. While replacing the pure glycerol by crude glycerol (60%), the media supported the cellular growth of st.BG10 (lac1), and produced 2343 ± 60 U/l of laccase after 48 h. The free cell biomass of st.BG10 (lac1) was recycled for 6times and produced > 8800U of laccase by using pure/crude glycerol media. Simultaneously, different matrices (agar–agar, calcium alginate, and polyacrylamide-gel) were tested to immobilize the st.BG10 (lac1) for laccase production, in which 6% calcium alginate immobilized cells to the bead size of 2.98 ± 0.07 mm were found to be stable, and recycled for 18 batches with 27,681–33926 U of laccase production from pure/crude glycerol based media. These findings would significantly assist to develop cost effective bioprocess for laccase production from crude glycerol (a waste product of biodiesel industry), and this is the first report on fungal laccase production by recyclable recombinant yeast using crude glycerol as substrate. Graphical Abstract: [Figure not available: see fulltext.]
dc.language	en
dc.publisher	Springer
dc.relation.ispartof	Waste and Biomass Valorization
dc.relation.isbasedon	10.1007/s12649-021-01661-1
dc.rights	info:eu-repo/semantics/closedAccess
dc.subject	0399 Other Chemical Sciences, 0904 Chemical Engineering, 0907 Environmental Engineering
dc.title	High Production of Trametes cinnabarina Laccase (lac1) by Suspended and Immobilized Cells of Recombinant Pichia pastoris from Crude Glycerol
dc.type	Journal Article
utslib.citation.volume	13
utslib.for	0399 Other Chemical Sciences
utslib.for	0904 Chemical Engineering
utslib.for	0907 Environmental Engineering
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Health
pubs.organisational-group	/University of Technology Sydney/Strength - CFI - Centre for Inflammation
utslib.copyright.status	closed_access	*
pubs.consider-herdc	false
dc.date.updated	2022-11-09T04:05:11Z
pubs.issue	4
pubs.publication-status	Published
pubs.volume	13
utslib.citation.issue	4

Abstract:

Laccases are green catalysts widely employed in various industrial, environmental, clinical and biotechnological applications. This study was aimed to develop the cost effective and efficient process for high level of fungal laccase production from crude glycerol. Initially, laccase gene (lac1) of Trametes cinnabarina was chemically synthesized, and integrated into Pichia pastoris strains (BG10, 11 and 16) by using pD915 vector which can facilitate the constitutive secretory expression of laccase (lac1). Among the recombinant P. pastoris strains developed, st.BG10 (wild type) was supported the high level of laccase production (2851 ± 72 U/l) from pure glycerol based media. While replacing the pure glycerol by crude glycerol (60%), the media supported the cellular growth of st.BG10 (lac1), and produced 2343 ± 60 U/l of laccase after 48 h. The free cell biomass of st.BG10 (lac1) was recycled for 6times and produced > 8800U of laccase by using pure/crude glycerol media. Simultaneously, different matrices (agar–agar, calcium alginate, and polyacrylamide-gel) were tested to immobilize the st.BG10 (lac1) for laccase production, in which 6% calcium alginate immobilized cells to the bead size of 2.98 ± 0.07 mm were found to be stable, and recycled for 18 batches with 27,681–33926 U of laccase production from pure/crude glycerol based media. These findings would significantly assist to develop cost effective bioprocess for laccase production from crude glycerol (a waste product of biodiesel industry), and this is the first report on fungal laccase production by recyclable recombinant yeast using crude glycerol as substrate. Graphical Abstract: [Figure not available: see fulltext.]

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