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Jina SON,Kei-Anne BARITUGO,Yu Jung SOHN,Kyoung Hee KANG,Hee Taek KIM,Jeong Chan JOO,Si Jae PARK 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10
Gamma-aminobutyrate (GABA) is a valuable amino-carboxylic acid that is used in food and pharmaceutical industry. Biosynthesis of GABA involves the decarboxylation of l-glutamate using glutamate decarboxylase (GAD). Thus, C. glutamicum with heterologous expression of GAD has been used for direct fermentative production of GABA from glucose. However, GABA production is not efficient since there is a significant difference between optimal pH for GAD activity (pH 4.0) and cell growth (pH 7.0). Thus, we aimed to develop recombinant C. glutamicum strains with expression of GAD from various microorganisms and compared their ability to produce GABA at near neutral pH. Here, we will present the development of metabolically engineered C. glutamicum strains for one-step fermentative GABA production from glucose at optimally balanced neutral pH.
Kim, Hee Taek,Khang, Tae Uk,Baritugo, Kei-Anne,Hyun, Sung Min,Kang, Kyoung Hee,Jung, Sol Hee,Song, Bong Keun,Park, Kyungmoon,Oh, Min-Kyu,Kim, Gi Bae,Kim, Hyun Uk,Lee, Sang Yup,Park, Si Jae,Joo, Jeong Elsevier 2019 Metabolic engineering Vol.51 No.-
<P><B>Abstract</B></P> <P> <I>Corynebacterium glutamicum</I> was metabolically engineered for the production of glutaric acid, a C5 dicarboxylic acid that can be used as platform building block chemical for nylons and plasticizers. <I>C. glutamicum gabT</I> and <I>gabD</I> genes and <I>Pseudomonas putida davT</I> and <I>davD</I> genes encoding 5-aminovalerate transaminase and glutarate semialdehyde dehydrogenase, respectively, were examined in <I>C. glutamicum</I> for the construction of a glutaric acid biosynthesis pathway along with <I>P. putida davB</I> and <I>davA</I> genes encoding lysine 2-monooxygenase and delta-aminovaleramidase, respectively. The glutaric acid biosynthesis pathway constructed in recombinant <I>C. glutamicum</I> was engineered by examining strong synthetic promoters P<SUB>H30</SUB> and P<SUB>H36</SUB>, <I>C. glutamicum</I> codon-optimized <I>davTDBA</I> genes, and modification of <I>davB</I> gene with an N-terminal His<SUB>6</SUB>-tag to improve the production of glutaric acid. It was found that use of N-terminal His<SUB>6</SUB>-tagged DavB was most suitable for the production of glutaric acid from glucose. Fed-batch fermentation using the final engineered <I>C. glutamicum</I> H30_GA<SUB>His</SUB> strain, expressing <I>davTDA</I> genes along with <I>davB</I> fused with His<SUB>6</SUB>-tag at N-terminus could produce 24.5 g/L of glutaric acid with low accumulation of <SMALL>L</SMALL>-lysine (1.7 g/L), wherein 5-AVA accumulation was not observed during fermentation.</P> <P><B>Highlights</B></P> <P> <UL> <LI> <I>C. glutamicum</I> strains were metabolically engineered for the production of glutaric acid. </LI> <LI> Optimization of glutaric acid biosynthesis pathway could enhance glutaric acid production. </LI> <LI> Recombinant <I>C. glutamicum</I> strains efficiently produced glutaric acid from glucose. </LI> <LI> Fed-batch culture of recombinant <I>C. glutamicum</I> H30_GA<SUB>His</SUB> supported high-level production of glutaric acid from glucose. </LI> </UL> </P>
Metabolic Engineering of Corynebacterium glutamicum for the Production of 5-Hydroxyvaleric Acid
Yu Jung SOHN,Minsoo KANG,Kei-Anne BARITUGO,Jina SON,Kyoung Hee KANG,Mi Hee RYU,Siseon LEE,Mingi SOHN,Ye Jean JUNG,Kyungmoon PARK,Si Jae PARK,Jeong Chan JOO,Hee Taek KIM 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.4