Synthesis of di-rhamnolipids by the avirulent, mono-rhamnolipid producing strain Pseudomonas aeruginosa ATCC 9027.

IF 2 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Biotechnology Letters Pub Date : 2024-12-01 Epub Date: 2024-09-03 DOI:10.1007/s10529-024-03527-7
Abigail González-Valdez, Paola G Vázquez-Bueno, Jessica Hernández-Pineda, Gloria Soberón-Chávez
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引用次数: 0

Abstract

To construct a derivative of the avirulent Pseudomonas aeruginosa ATCC 9027 that produces high levels of di-rhamnolipid, that has better physico-chemical characteristics for biotechnological applications than mono-rhamnolipid, which is the sole type produced by ATCC 9027. We used plasmids expressing the rhlC gene, which encodes for rhamnosyl transferase II that transforms mono- to di-rhamnolipids under different promoters and in combination with the gene coding for the RhlR quorum sensing regulator, or the mono-rhamnolipid biosynthetic rhlAB operon. The plasmids tested carrying the rhlC gene under the lac promoter were plasmid prhlC and prhlRC, while prhlAB-R-C expressed this gene from the rhlA promoter, forming part of the artificially constructed rhlAB-R-C operon. We measured rhamnolipds concentrations using the orcinol method and determined the proportion of mono-rhamnolipids and di-rhamnolipids by UPLC/MS/MS. We found that the expression of rhlC in P. aeruginosa ATCC 9027 caused the production of di-rhamnolipids and that the derivative carrying plasmid prhlAB-R-C gives the best results considering total rhamnolipids and a higher proportion of di-rhamnolipids. A P. aeruginosa ATCC 9027 derivative with increased di-rhamnolipids production was developed by expressing plasmid prhlAB-R-C, that produces similar rhamnolipids levels as PAO1 type-strain and presented a higher proportion of di-rhamnolipids than this type-strain.

Abstract Image

产生单鼠李糖脂的无毒铜绿假单胞菌 ATCC 9027 株合成二鼠李糖脂。
为了构建一种无毒铜绿假单胞菌 ATCC 9027 的衍生物,该衍生物可产生大量二鼠李糖脂,与 ATCC 9027 唯一产生的单鼠李糖脂相比,二鼠李糖脂具有更好的理化特性,可用于生物技术应用。我们使用了表达 rhlC 基因或单鼠李糖脂生物合成 rhlAB 操作子的质粒,rhlC 基因编码鼠李糖基转移酶 II,该酶 II 可在不同的启动子下将单鼠李糖脂转化为双鼠李糖脂,并与编码 RhlR 法定量感应调节器的基因结合使用。在 lac 启动子下携带 rhlC 基因的质粒是质粒 prhlC 和 prhlRC,而 prhlAB-R-C 则在 rhlA 启动子下表达该基因,是人工构建的 rhlAB-R-C 操作子的一部分。我们使用奥克诺尔法测定了鼠李糖的浓度,并通过 UPLC/MS/MS 测定了单鼠李糖和二鼠李糖的比例。我们发现,rhlC 在铜绿微囊藻 ATCC 9027 中的表达会导致二鼠李糖脂的产生,而携带质粒 prhlAB-R-C 的衍生物在考虑总鼠李糖脂和较高比例的二鼠李糖脂时结果最好。通过表达质粒prhlAB-R-C,开发出一种铜绿微囊藻ATCC 9027衍生物,该衍生物具有更高的二鼠李糖脂产量,其鼠李糖脂产量与PAO1型菌株相似,但二鼠李糖脂比例高于该型菌株。
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来源期刊
Biotechnology Letters
Biotechnology Letters 工程技术-生物工程与应用微生物
CiteScore
5.90
自引率
3.70%
发文量
108
审稿时长
1.2 months
期刊介绍: Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.
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