Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus

Summary Salinomycin, an FDA‐approved polyketide drug, was recently identified as a promising anti‐tumour and anti‐viral lead compound. It is produced by Streptomyces albus, and the biosynthetic gene cluster (sal) spans over 100 kb. The genetic manipulation of large polyketide gene clusters is challe...

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Autores principales:	Dong Li, Yuqing Tian, Xiang Liu, Wenxi Wang, Yue Li, Huarong Tan, Jihui Zhang
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/8d7d8086f5dc4ce2807bd5fb8a825ef6
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spelling	oai:doaj.org-article:8d7d8086f5dc4ce2807bd5fb8a825ef62021-11-18T15:39:52ZReconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus1751-791510.1111/1751-7915.13686https://doaj.org/article/8d7d8086f5dc4ce2807bd5fb8a825ef62021-11-01T00:00:00Zhttps://doi.org/10.1111/1751-7915.13686https://doaj.org/toc/1751-7915Summary Salinomycin, an FDA‐approved polyketide drug, was recently identified as a promising anti‐tumour and anti‐viral lead compound. It is produced by Streptomyces albus, and the biosynthetic gene cluster (sal) spans over 100 kb. The genetic manipulation of large polyketide gene clusters is challenging, and approaches delivering reliable efficiency and accuracy are desired. Herein, a delicate strategy to enhance salinomycin production was devised and evaluated. We reconstructed a minimized sal gene cluster (mini‐cluster) on pSET152 including key genes responsible for tailoring modification, antibiotic resistance, positive regulation and precursor supply. These genes were overexpressed under the control of constitutive promoter PkasO* or Pneo. The pks operon was not included in the mini‐cluster, but it was upregulated by SalJ activation. After the plasmid pSET152::mini‐cluster was introduced into the wild‐type strain and a chassis host strain obtained by ribosome engineering, salinomycin production was increased to 2.3‐fold and 5.1‐fold compared with that of the wild‐type strain respectively. Intriguingly, mini‐cluster introduction resulted in much higher production than overexpression of the whole sal gene cluster. The findings demonstrated that reconstitution of sal mini‐cluster combined with ribosome engineering is an efficient novel approach and may be extended to other large polyketide biosynthesis.Dong LiYuqing TianXiang LiuWenxi WangYue LiHuarong TanJihui ZhangWileyarticleBiotechnologyTP248.13-248.65ENMicrobial Biotechnology, Vol 14, Iss 6, Pp 2356-2368 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biotechnology TP248.13-248.65
spellingShingle	Biotechnology TP248.13-248.65 Dong Li Yuqing Tian Xiang Liu Wenxi Wang Yue Li Huarong Tan Jihui Zhang Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
description	Summary Salinomycin, an FDA‐approved polyketide drug, was recently identified as a promising anti‐tumour and anti‐viral lead compound. It is produced by Streptomyces albus, and the biosynthetic gene cluster (sal) spans over 100 kb. The genetic manipulation of large polyketide gene clusters is challenging, and approaches delivering reliable efficiency and accuracy are desired. Herein, a delicate strategy to enhance salinomycin production was devised and evaluated. We reconstructed a minimized sal gene cluster (mini‐cluster) on pSET152 including key genes responsible for tailoring modification, antibiotic resistance, positive regulation and precursor supply. These genes were overexpressed under the control of constitutive promoter PkasO* or Pneo. The pks operon was not included in the mini‐cluster, but it was upregulated by SalJ activation. After the plasmid pSET152::mini‐cluster was introduced into the wild‐type strain and a chassis host strain obtained by ribosome engineering, salinomycin production was increased to 2.3‐fold and 5.1‐fold compared with that of the wild‐type strain respectively. Intriguingly, mini‐cluster introduction resulted in much higher production than overexpression of the whole sal gene cluster. The findings demonstrated that reconstitution of sal mini‐cluster combined with ribosome engineering is an efficient novel approach and may be extended to other large polyketide biosynthesis.
format	article
author	Dong Li Yuqing Tian Xiang Liu Wenxi Wang Yue Li Huarong Tan Jihui Zhang
author_facet	Dong Li Yuqing Tian Xiang Liu Wenxi Wang Yue Li Huarong Tan Jihui Zhang
author_sort	Dong Li
title	Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
title_short	Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
title_full	Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
title_fullStr	Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
title_full_unstemmed	Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus
title_sort	reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in streptomyces albus
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/8d7d8086f5dc4ce2807bd5fb8a825ef6
work_keys_str_mv	AT dongli reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT yuqingtian reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT xiangliu reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT wenxiwang reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT yueli reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT huarongtan reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus AT jihuizhang reconstitutionofaminigeneclustercombinedwithribosomeengineeringledtoeffectiveenhancementofsalinomycinproductioninstreptomycesalbus
_version_	1718420796424060928

Reconstitution of a mini‐gene cluster combined with ribosome engineering led to effective enhancement of salinomycin production in Streptomyces albus

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