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Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application

BACKGROUND: A lot of kitchen waste oil is produced every day worldwide, leading to serious environmental pollution. As one of the environmental protection methods, microorganisms are widely used treating of various wastes. Lipase, as one of the cleaning agents can effectively degrade kitchen waste o...

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Autores principales: Duan, Xinran, Zhai, Wei, Li, Xintian, Wu, Sicheng, Wang, Ye, Wang, Lixia, Basang, Wangdui, Zhu, Yanbin, Gao, Yunhang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10625193/
https://www.ncbi.nlm.nih.gov/pubmed/37924095
http://dx.doi.org/10.1186/s12896-023-00818-1
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author Duan, Xinran
Zhai, Wei
Li, Xintian
Wu, Sicheng
Wang, Ye
Wang, Lixia
Basang, Wangdui
Zhu, Yanbin
Gao, Yunhang
author_facet Duan, Xinran
Zhai, Wei
Li, Xintian
Wu, Sicheng
Wang, Ye
Wang, Lixia
Basang, Wangdui
Zhu, Yanbin
Gao, Yunhang
author_sort Duan, Xinran
collection PubMed
description BACKGROUND: A lot of kitchen waste oil is produced every day worldwide, leading to serious environmental pollution. As one of the environmental protection methods, microorganisms are widely used treating of various wastes. Lipase, as one of the cleaning agents can effectively degrade kitchen waste oil. The composting process of pig carcasses produces many lipase producing microorganisms, rendering compost products an excellent source for isolating lipase producing microorganisms. To our knowledge, there are no reports isolating of lipase producing strains from the high temperature phase of pig carcass compost. METHODOLOGY: Lipase producing strains were isolated using a triglyceride medium and identified by 16S rRNA gene sequencing. The optimal fermentation conditions for maximum lipase yield were gradually optimized by single-factor tests. The extracellular lipase was purified by ammonium sulfate precipitation and Sephadex G-75 gel isolation chromatography. Amino acid sequence analysis, structure prediction, and molecular docking of the purified protein were performed. The pure lipase's enzymatic properties and application potential were evaluated by characterizing its biochemical properties. RESULTS: In this study, a lipase producing strain of Bacillus sp. ZF2 was isolated from pig carcass compost products, the optimal fermentation conditions of lipase: sucrose 3 g/L, ammonium sulfate 7 g/L, Mn(2+) 1.0 mmol/L, initial pH 6, inoculum 5%, temperature 25 ℃, and fermentation time 48 h. After purification, the specific activity of the purified lipase reached 317.59 U/mg, a 9.78-fold improvement. Lipase had the highest similarity to the GH family 46 chitosanase and molecular docking showed that lipase binds to fat via two hydrogen bonds at Gln146 (A) and Glu203 (A). Under different conditions (temperature, metal ions, organic solvents, and surfactants), lipase can maintain enzymatic activity. Under different types of kitchen oils, lipase has low activity only for ‘chicken oil’, in treating other substrates, the enzyme activity can exceed 50%. CONCLUSIONS: This study reveals the potential of lipase for waste oil removal, and future research will be devoted to the application of lipase. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-023-00818-1.
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spelling pubmed-106251932023-11-05 Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application Duan, Xinran Zhai, Wei Li, Xintian Wu, Sicheng Wang, Ye Wang, Lixia Basang, Wangdui Zhu, Yanbin Gao, Yunhang BMC Biotechnol Research BACKGROUND: A lot of kitchen waste oil is produced every day worldwide, leading to serious environmental pollution. As one of the environmental protection methods, microorganisms are widely used treating of various wastes. Lipase, as one of the cleaning agents can effectively degrade kitchen waste oil. The composting process of pig carcasses produces many lipase producing microorganisms, rendering compost products an excellent source for isolating lipase producing microorganisms. To our knowledge, there are no reports isolating of lipase producing strains from the high temperature phase of pig carcass compost. METHODOLOGY: Lipase producing strains were isolated using a triglyceride medium and identified by 16S rRNA gene sequencing. The optimal fermentation conditions for maximum lipase yield were gradually optimized by single-factor tests. The extracellular lipase was purified by ammonium sulfate precipitation and Sephadex G-75 gel isolation chromatography. Amino acid sequence analysis, structure prediction, and molecular docking of the purified protein were performed. The pure lipase's enzymatic properties and application potential were evaluated by characterizing its biochemical properties. RESULTS: In this study, a lipase producing strain of Bacillus sp. ZF2 was isolated from pig carcass compost products, the optimal fermentation conditions of lipase: sucrose 3 g/L, ammonium sulfate 7 g/L, Mn(2+) 1.0 mmol/L, initial pH 6, inoculum 5%, temperature 25 ℃, and fermentation time 48 h. After purification, the specific activity of the purified lipase reached 317.59 U/mg, a 9.78-fold improvement. Lipase had the highest similarity to the GH family 46 chitosanase and molecular docking showed that lipase binds to fat via two hydrogen bonds at Gln146 (A) and Glu203 (A). Under different conditions (temperature, metal ions, organic solvents, and surfactants), lipase can maintain enzymatic activity. Under different types of kitchen oils, lipase has low activity only for ‘chicken oil’, in treating other substrates, the enzyme activity can exceed 50%. CONCLUSIONS: This study reveals the potential of lipase for waste oil removal, and future research will be devoted to the application of lipase. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12896-023-00818-1. BioMed Central 2023-11-03 /pmc/articles/PMC10625193/ /pubmed/37924095 http://dx.doi.org/10.1186/s12896-023-00818-1 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Duan, Xinran
Zhai, Wei
Li, Xintian
Wu, Sicheng
Wang, Ye
Wang, Lixia
Basang, Wangdui
Zhu, Yanbin
Gao, Yunhang
Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title_full Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title_fullStr Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title_full_unstemmed Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title_short Preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
title_sort preparation, purification, and biochemical of fat-degrading bacterial enzymes from pig carcass compost and its application
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10625193/
https://www.ncbi.nlm.nih.gov/pubmed/37924095
http://dx.doi.org/10.1186/s12896-023-00818-1
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