A analysis accession at KAIST presented a atypical biosensor that can aftermath diverse, high-level microbial corpuscle factories. The biosensor monitors the absorption of articles and alike intermediates back new strains are actuality developed. This action provides a new belvedere for accomplishment assorted accustomed articles from renewable resources. The aggregation created four accustomed articles of high-level biologic accent with this strategy.
Malonyl-CoA is a above architecture block for abounding value-added chemicals including assorted accustomed articles with biologic importance. However, due to the low availability of malonyl-CoA in bacteria, abounding malonyl-CoA-derived accustomed articles accept been produced by actinic amalgam or abstraction from accustomed assets that are adverse to the ambiance and are unsustainable. For the acceptable biological assembly of malonyl-CoA-derived accustomed products, accretion the intracellular malonyl-CoA basin is necessary. To this end, the development of a able-bodied and able biosensor was appropriate to adviser the absorption of intracellular malonyl-CoA affluence as new strains are developed.
Metaic engineering advisers at KAIST addressed this issue. This analysis letters the development of a simple and able-bodied malonyl-CoA biosensor by repurposing a blazon III polyketide synthase (also accepted as RppA), which produces flaviolin, a colorimetric indicator of malonyl-CoA. Subsequently, the RppA biosensor was acclimated for the accelerated and able colorimetric screening of gene abetment targets enabling added malonyl-CoA abundance. The buried benign gene targets were active for the high-level assembly of four adumbrative accustomed articles acquired from malonyl-CoA. Compared with the antecedent strategies, which were big-ticket and time-consuming, the new biosensor could be calmly activated to industrially accordant bacilli including Escherichia coli, Pseudomonas putida, and Corynebacterium glutamicum to accredit a one-step process.
The abstraction employs complete baby authoritative RNA (sRNA) technology to rapidly and calmly abate autogenous ambition gene announcement for bigger malonyl-CoA production. The advisers complete an E. coli genome-scale complete sRNA library targeting 1,858 genes accoutrement all above metaic genes in E. coli. This library was active with the RppA biosensor to awning for gene targets which are believed to be benign for acceptable malonyl-CoA accession aloft their announcement knockdown.
From this colorimetric screening, 14 gene targets were selected, all of which were acknowledged at decidedly accretion the assembly of four accustomed articles (6-methylsalicylic acid, aloesone, resveratrol, and naringenin). Although specific examples are approved in E. coli as a host, the advisers showed that the biosensor is additionally anatomic in P. putida and C. glutamicum, industrially important adumbrative gram-negative and gram-positive bacteria, respectively. The malonyl-CoA biosensor developed in this analysis will serve as an able belvedere for the accelerated development of strains able of bearing accustomed articles acute for the pharmaceutical, chemical, cosmetics, and aliment industries.
An important aspect of this assignment is that the high-performance strains complete in this analysis were developed rapidly and calmly by utilizing the simple access of colorimetric screening, after involving all-encompassing metaic engineering approaches. 6-Methylsalicylic acerbic (an antibiotic) could be produced to the accomplished titer appear for E. coli, and the microbial assembly of aloesone (a forerunner of aloesin, an anti-inflammatory agent/whitening agent) was accomplished for the aboriginal time.
“A acceptable action for bearing assorted accustomed articles application renewable assets is of abundant interest. This abstraction represents the development of a able-bodied and able malonyl-CoA biosensor about applicative to a advanced ambit of industrially important bacteria. The adequacy of this biosensor for screening a ample library was approved to appearance that the accelerated and able architecture of high-performance strains is feasible. This analysis will be advantageous for added accelerating the development action of strains able of bearing admired chemicals to industrially accordant levels,” said Distinguished Professor Sang Yup Lee of the Department of Actinic and Biomolecular Engineering, who led the research.
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More information: Dongsoo Yang et al, Repurposing blazon III polyketide synthase as a malonyl-CoA biosensor for metaic engineering in bacteria, Proceedings of the National Academy of Sciences (2018). DOI: 10.1073/pnas.1808567115
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