Gene editing technology combined with response surface optimization to improve the synthesis ability of lycopene in Pantoea dispersa MSC14.

IF 3.2 3区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of Applied Microbiology Pub Date : 2024-11-04 DOI:10.1093/jambio/lxae272
La Lai, Run Xin, Tangbing Cui
{"title":"Gene editing technology combined with response surface optimization to improve the synthesis ability of lycopene in Pantoea dispersa MSC14.","authors":"La Lai, Run Xin, Tangbing Cui","doi":"10.1093/jambio/lxae272","DOIUrl":null,"url":null,"abstract":"<p><strong>Aim: </strong>The aim of this study is to engineer Pantoea dispersa MSC14 into a strain capable of producing lycopene and to enhance its lycopene content.</p><p><strong>Methods and results: </strong>Our laboratory isolated the strain P. dispersa MSC14 from petroleum-contaminated soil in a mining area. Whole-genome sequencing confirmed the existence of a carotenoid synthesis pathway in this strain. This study employed an optimized CRISPR/Cas9 system to perform a traceless gene knockout of the lycopene cyclase gene crtY and to overexpress the octahydrolycopene dehydrogenase gene crtI in the P. dispersa MSC14. This strategic genetic modification successfully constructed the lycopene-producing strain MSC14-LY, which exhibited a notable lycopene content with a biomass productivity of 553 μg of lycopene per gram dry cell weight (DCW). Additionally, the components of the lycopene fermentation medium were optimized using Plackett-Burman design and response surface methodology. The average lycopene content was increased to 5.13 mg g -1 DCW in the optimized LY fermentation medium. Through genetic engineering, P. dispersa MSC14 was transformed into a strain capable of producing lycopene, achieving a yield of 5.13 mg g-1 DCW after medium optimization.</p><p><strong>Conclusions: </strong>Genetic engineering successfully transformed P. dispersa MSC14 into a strain capable of producing lycopene, achieving a yield of 5.13 mg g-1 DCW after medium optimization.</p>","PeriodicalId":15036,"journal":{"name":"Journal of Applied Microbiology","volume":" ","pages":""},"PeriodicalIF":3.2000,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Applied Microbiology","FirstCategoryId":"88","ListUrlMain":"https://doi.org/10.1093/jambio/lxae272","RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

Abstract

Aim: The aim of this study is to engineer Pantoea dispersa MSC14 into a strain capable of producing lycopene and to enhance its lycopene content.

Methods and results: Our laboratory isolated the strain P. dispersa MSC14 from petroleum-contaminated soil in a mining area. Whole-genome sequencing confirmed the existence of a carotenoid synthesis pathway in this strain. This study employed an optimized CRISPR/Cas9 system to perform a traceless gene knockout of the lycopene cyclase gene crtY and to overexpress the octahydrolycopene dehydrogenase gene crtI in the P. dispersa MSC14. This strategic genetic modification successfully constructed the lycopene-producing strain MSC14-LY, which exhibited a notable lycopene content with a biomass productivity of 553 μg of lycopene per gram dry cell weight (DCW). Additionally, the components of the lycopene fermentation medium were optimized using Plackett-Burman design and response surface methodology. The average lycopene content was increased to 5.13 mg g -1 DCW in the optimized LY fermentation medium. Through genetic engineering, P. dispersa MSC14 was transformed into a strain capable of producing lycopene, achieving a yield of 5.13 mg g-1 DCW after medium optimization.

Conclusions: Genetic engineering successfully transformed P. dispersa MSC14 into a strain capable of producing lycopene, achieving a yield of 5.13 mg g-1 DCW after medium optimization.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
基因编辑技术与响应面优化相结合,提高散囊菌 MSC14 的番茄红素合成能力。
目的:本研究的目的是将散囊菌 MSC14 改造成能够生产番茄红素的菌株,并提高其番茄红素含量:我们的实验室从矿区受石油污染的土壤中分离出了散囊菌MSC14菌株。全基因组测序证实该菌株存在类胡萝卜素合成途径。本研究利用优化的 CRISPR/Cas9 系统对 P. dispersa MSC14 中的番茄红素环化酶基因 crtY 进行无痕基因敲除,并过表达八氢番茄红素脱氢酶基因 crtI。这种战略性基因改造成功构建了番茄红素生产菌株 MSC14-LY,其番茄红素含量显著提高,生物量生产率达到每克干细胞重量(DCW)553 微克番茄红素。此外,还利用普拉克特-伯曼(PB)设计和响应面法(RSM)对番茄红素发酵培养基的成分进行了优化。在优化的 LY 发酵培养基中,番茄红素的平均含量提高到 5.13 mg g -1DCW 。通过基因工程将 P. dispersa MSC14 转化为能够生产番茄红素的菌株,在培养基优化后,产量达到 5.13 mg g-1 DCW:结论:基因工程成功地将 P. dispersa MSC14 转化为能够生产番茄红素的菌株,经过培养基优化后,产量达到 5.13 mg g-1 DCW。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Journal of Applied Microbiology
Journal of Applied Microbiology 生物-生物工程与应用微生物
CiteScore
7.30
自引率
2.50%
发文量
427
审稿时长
2.7 months
期刊介绍: Journal of & Letters in Applied Microbiology are two of the flagship research journals of the Society for Applied Microbiology (SfAM). For more than 75 years they have been publishing top quality research and reviews in the broad field of applied microbiology. The journals are provided to all SfAM members as well as having a global online readership totalling more than 500,000 downloads per year in more than 200 countries. Submitting authors can expect fast decision and publication times, averaging 33 days to first decision and 34 days from acceptance to online publication. There are no page charges.
期刊最新文献
Efficacy of photoClO2 against two human norovirus surrogates and Clostridioides difficile endospores on stainless steel and nylon carpet. Influence of feeding black soldier fly (Hermetia illucens), cricket (Gryllodes sigillatus), and superworm (Zophobas morio) on the gut microbiota of rainbow trout (Oncorhynchus mykiss). Occurrence of Salmonella enterica in faecal sludge from Nigeria and genetic relatedness with strains associated with human infections in Africa. Epidemiology and pathogen characteristics of infections following solid organ transplantation. Glucose-1-phosphate thymidylyltransferase promotes the production of 3-O-α-mycarosylerythronolide B in Streptomyces coelicolor.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1