Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) co-produced with L-isoleucine in Corynebacterium glutamicum WM001.

Publisher: BioMed Central Country of Publication: England NLM ID: 101139812 Publication Model: Electronic Cited Medium: Internet ISSN: 1475-2859 (Electronic) Linking ISSN: 14752859 NLM ISO Abbreviation: Microb Cell Fact Subsets: MEDLINE

Original Publication: London : BioMed Central, [2002-

Acyl Coenzyme A/*metabolism ; Corynebacterium glutamicum/*metabolism ; Hydroxybutyrates/*metabolism ; Isoleucine/*metabolism

Background: Co-production of polyhydroxyalkanoate (PHA) and amino acids makes bacteria effective microbial cell factories by secreting amino acids outside while accumulating PHA granules inside. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is one of the PHAs with biocompatibility and fine mechanical properties, but its production is limited by the low level of intracellular propionyl-CoA.
Results: L-Isoleucine producing Corynebacterium glutamicum strain WM001 were analyzed by genome and transcriptome sequencing. The results showed that the most over-expressed genes in WM001 are relevant not only to L-isoleucine production but also to propionyl-CoA accumulation. Compared to the wild-type C. glutamicum ATCC13869, the transcriptional levels of the genes prpC2, prpD2, and prpB2, which are key genes relevant to propionyl-CoA accumulation, increased 2 ^6.7 , 2 ^5.8 , and 2 ^8.4 -folds in WM001, respectively; and the intracellular level of propionyl-CoA increased 16.9-fold in WM001. When the gene cluster phaCAB for PHA biosynthesis was introduced into WM001, the recombinant strain WM001/pDXW-8-phaCAB produced 15.0 g/L PHBV with high percentage of 3-hydroxyvalerate as well as 29.8 g/L L-isoleucine after fed-batch fermentation. The maximum 3-hydroxyvalerate fraction in PHBV produced by WM001/pDXW-8-phaCAB using glucose as the sole carbon source could reach 72.5%, which is the highest reported so far.
Conclusions: Genome and transcriptome analysis showed that C. glutamicum WM001 has potential to accumulate L-isoleucine and propionyl-CoA pool. This was experimentally confirmed by introducing the phaCAB gene cluster into WM001. The recombinant strain WM001/pDXW-8-phaCAB produced high levels of PHBV with high 3-hydroxyvalerate fraction as well as L-isoleucine. Because of its high level of intracellular propionyl-CoA pool, WM001 might be used for producing other propionyl-CoA derivatives.

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LITE2018-10 National First-class Discipline Program of Light Industry Technology and Engineering

Keywords: 3HV fraction; Corynebacterium glutamicum; L-Isoleucine; PHBV; Propionyl-CoA

0 (Acyl Coenzyme A)
0 (Hydroxybutyrates)
04Y7590D77 (Isoleucine)
317-66-8 (propionyl-coenzyme A)

Date Created: 20180617 Date Completed: 20181005 Latest Revision: 20181114

20240829

PMC6004086

10.1186/s12934-018-0942-7

29907151