Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains

Abstract Bioethanol produced by fermentative microorganisms is regarded as an alternative to fossil fuel. Bioethanol to be used as a viable energy source must be produced cost-effectively by removing expense-intensive steps such as the enzymatic hydrolysis of substrate. Consolidated bioprocessing (C...

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Autores principales:	Fatemeh Maleki, Mohammad Changizian, Narges Zolfaghari, Sarah Rajaei, Kambiz Akbari Noghabi, Hossein Shahbani Zahiri
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/68ff566b6f8d4cc7bd13ce35d77b1783
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spelling	oai:doaj.org-article:68ff566b6f8d4cc7bd13ce35d77b17832021-12-02T16:31:57ZConsolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains10.1038/s41598-021-92627-92045-2322https://doaj.org/article/68ff566b6f8d4cc7bd13ce35d77b17832021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92627-9https://doaj.org/toc/2045-2322Abstract Bioethanol produced by fermentative microorganisms is regarded as an alternative to fossil fuel. Bioethanol to be used as a viable energy source must be produced cost-effectively by removing expense-intensive steps such as the enzymatic hydrolysis of substrate. Consolidated bioprocessing (CBP) is believed to be a practical solution combining saccharification and fermentation in a single step catalyzed by a microorganism. Bacillus subtills with innate ability to grow on a diversity of carbohydrates seems promising for affordable CBP bioethanol production using renewable plant biomass and wastes. In this study, the genes encoding alcohol dehydrogenase from Z. mobilis (adh Z ) and S. cerevisiae (adh S ) were each used with Z. mobilis pyruvate decarboxylase gene (pdc Z ) to create ethanologenic operons in a lactate-deficient (Δldh) B. subtilis resulting in NZ and NZS strains, respectively. The S. cerevisiae adh S caused significantly more ethanol production by NZS and therefore was used to make two other operons including one with double copies of both pdc Z and adh S and the other with a single pdc Z but double adh S genes expressed in N(ZS)2 and NZS2 strains, respectively. In addition, two fusion genes were constructed with pdc Z and adh S in alternate orientations and used for ethanol production by the harboring strains namely NZ:S and NS:Z, respectively. While the increase of gene dosage was not associated with elevated carbon flow for ethanol production, the fusion gene adh S:pdc Z resulted in a more than two times increase of productivity by strain NS:Z as compared with NZS during 48 h fermentation. The CBP ethanol production by NZS and NS:Z using potatoes resulted in 16.3 g/L and 21.5 g/L ethanol during 96 h fermentation, respectively. For the first time in this study, B. subtilis was successfully used for CBP ethanol production with S. cerevisiae alcohol dehydrogenase. The results of the study provide insights on the potentials of B. subtilis for affordable bioethanol production from inexpensive plant biomass and wastes. However, the potentials need to be improved by metabolic and process engineering for higher yields of ethanol production and plant biomass utilization.Fatemeh MalekiMohammad ChangizianNarges ZolfaghariSarah RajaeiKambiz Akbari NoghabiHossein Shahbani ZahiriNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Fatemeh Maleki Mohammad Changizian Narges Zolfaghari Sarah Rajaei Kambiz Akbari Noghabi Hossein Shahbani Zahiri Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
description	Abstract Bioethanol produced by fermentative microorganisms is regarded as an alternative to fossil fuel. Bioethanol to be used as a viable energy source must be produced cost-effectively by removing expense-intensive steps such as the enzymatic hydrolysis of substrate. Consolidated bioprocessing (CBP) is believed to be a practical solution combining saccharification and fermentation in a single step catalyzed by a microorganism. Bacillus subtills with innate ability to grow on a diversity of carbohydrates seems promising for affordable CBP bioethanol production using renewable plant biomass and wastes. In this study, the genes encoding alcohol dehydrogenase from Z. mobilis (adh Z ) and S. cerevisiae (adh S ) were each used with Z. mobilis pyruvate decarboxylase gene (pdc Z ) to create ethanologenic operons in a lactate-deficient (Δldh) B. subtilis resulting in NZ and NZS strains, respectively. The S. cerevisiae adh S caused significantly more ethanol production by NZS and therefore was used to make two other operons including one with double copies of both pdc Z and adh S and the other with a single pdc Z but double adh S genes expressed in N(ZS)2 and NZS2 strains, respectively. In addition, two fusion genes were constructed with pdc Z and adh S in alternate orientations and used for ethanol production by the harboring strains namely NZ:S and NS:Z, respectively. While the increase of gene dosage was not associated with elevated carbon flow for ethanol production, the fusion gene adh S:pdc Z resulted in a more than two times increase of productivity by strain NS:Z as compared with NZS during 48 h fermentation. The CBP ethanol production by NZS and NS:Z using potatoes resulted in 16.3 g/L and 21.5 g/L ethanol during 96 h fermentation, respectively. For the first time in this study, B. subtilis was successfully used for CBP ethanol production with S. cerevisiae alcohol dehydrogenase. The results of the study provide insights on the potentials of B. subtilis for affordable bioethanol production from inexpensive plant biomass and wastes. However, the potentials need to be improved by metabolic and process engineering for higher yields of ethanol production and plant biomass utilization.
format	article
author	Fatemeh Maleki Mohammad Changizian Narges Zolfaghari Sarah Rajaei Kambiz Akbari Noghabi Hossein Shahbani Zahiri
author_facet	Fatemeh Maleki Mohammad Changizian Narges Zolfaghari Sarah Rajaei Kambiz Akbari Noghabi Hossein Shahbani Zahiri
author_sort	Fatemeh Maleki
title	Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
title_short	Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
title_full	Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
title_fullStr	Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
title_full_unstemmed	Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains
title_sort	consolidated bioprocessing for bioethanol production by metabolically engineered bacillus subtilis strains
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/68ff566b6f8d4cc7bd13ce35d77b1783
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Consolidated bioprocessing for bioethanol production by metabolically engineered Bacillus subtilis strains

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