Microbial secretion of lactate-enriched oligomers for efficient conversion into lactide: A biological shortcut to polylactide

Utsunomia, Camila; Matsumoto, Ken’ichiro; Date, Sakiko; Hori, Chiaki; Taguchi, Seiichi

doi:10.1016/j.jbiosc.2017.03.002

Cited by 13 publications

(6 citation statements)

References 26 publications

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“…As mentioned in the introduction, 3HBOs without any modification are distributed as attractive physiologically active substances at various sites. In previous studies, lactate‐based oligomers consisted of two monomer units, lactate and 3HB, at different monomeric compositions . To utilize the secreted oligomers as industrial products, uniform oligomers (monomer composition, end structure, and molecular weight) are desirable.…”

Section: Discussionmentioning

confidence: 99%

“…first reported the secretory production of low‐molecular‐weight lactate‐based polymers (or oligomers) consisting of D‐lactate and ( R )‐3HB in a recombinant E. coli system using lactate‐polymerizing enzyme (LPE) . The secretion efficiency of lactate‐based oligomers was remarkably facilitated by an external addition of chain transfer (CT) agents such as diethylene glycol (DEG) . In addition, transporter‐dependent secretion was observed for the outer and inner membranes of E. coli .…”

Section: Introductionmentioning

confidence: 99%

“…In addition, transporter‐dependent secretion was observed for the outer and inner membranes of E. coli . Notably, the direct secretion of lactate‐based oligomers enabled us to establish a shortcut for producing poly(lactide) (PLA) via the ring‐opening polymerization of lactide converted from the lactate‐based oligomers . Secreted lactate‐based oligomers end‐capped with DEG were also functionally used to synthesize poly(ester‐urethane) .…”

Section: Introductionmentioning

confidence: 99%

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Microbial Secretion Platform for 3‐Hydroxybutyrate Oligomer and Its End‐Capped Forms Using Chain Transfer Reaction‐Mediated Polyhydroxyalkanoate Synthases

Miyahara

Hiroe

Tsuge

et al. 2019

Biotechnology Journal

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The biodegradable polyester 3‐hydroxybutyrate (3HB) polymer [P(3HB)] is intracellularly synthesized and accumulated in recombinant Escherichia coli. In this study, native polyhydroxyalkanoate (PHA) synthases are used to attempt to microbially secrete 3HB homo‐oligomers (3HBOs), which are widely distributed in nature as physiologically active substances. High secretory production is observed, especially for the two PHA synthases from Aeromonas caviae and Bacillus cereus YB4. Surprisingly, an ethyl ester at the carboxy terminus (ethyl ester form) of 3HBOs is identified for most of the PHA synthases tested. Next, 3HBOs with a functional carboxyl group (carboxyl form of 3HBO) are obtained by using the alcohol dehydrogenase gene (adhE)‐deficient mutant strain, suggesting that the endogenous ethanol produced in E. coli acts as a chain transfer (CT) agent in the generation of 3HBOs. Furthermore, an in vitro polymerization assay reveals that CT agents such as ethanol and free 3HB are involved in the generation of ethyl ester and carboxyl form of 3HBO, respectively. The microbial platform established herein allows the secretion of 3HBOs with desirable end structures by supplementation with various CT agents. The obtained 3HBOs and their end‐capped forms may be used as physiologically active substances and building blocks for polymeric materials.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Microbial Secretion Platform for 3‐Hydroxybutyrate Oligomer and Its End‐Capped Forms Using Chain Transfer Reaction‐Mediated Polyhydroxyalkanoate Synthases

Miyahara

Hiroe

Tsuge

et al. 2019

Biotechnology Journal

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“…In recent studies, the secretory production of low-molecular-weight D-lactate-based polymers and P(3HB) polymers, referred to as D-lactate-based oligomers (D-LAOs) and 3HB oligomers (3HBOs), respectively, was demonstrated using Escherichia coli heterologously expressing PHA synthases [13][14][15][16][17][18]. Unlike polymer production in cells, secretory production is a continuous process because of the lack of a cell volume limitation.…”

Section: Introductionmentioning

confidence: 99%

Enhanced Production of (R)-3-Hydroxybutyrate Oligomers by Coexpression of Molecular Chaperones in Recombinant Escherichia coli Harboring a Polyhydroxyalkanoate Synthase Derived from Bacillus cereus YB-4

et al. 2022

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The biodegradable polyester poly-(R)-3-hydroxybutyrate [P(3HB)] is synthesized by a polymerizing enzyme called polyhydroxyalkanoate (PHA) synthase and accumulates in a wide variety of bacterial cells. Recently, we demonstrated the secretory production of a (R)-3HB oligomer (3HBO), a low-molecular-weight P(3HB), by using recombinant Escherichia coli expressing PHA synthases. The 3HBO has potential value as an antibacterial substance and as a building block for various polymers. In this study, to construct an efficient 3HBO production system, the coexpression of molecular chaperones and a PHA synthase derived from Bacillus cereus YB-4 (PhaRCYB4) was examined. First, genes encoding enzymes related to 3HBO biosynthesis (phaRCYB4, phaA and phaB derived from Ralstonia eutropha H16) and two types of molecular chaperones (groEL, groES, and tig) were introduced into the E. coli strains BW25113 and BW25113ΔadhE. As a result, coexpression of the chaperones promoted the enzyme activity of PHA synthase (approximately 2–3-fold) and 3HBO production (approximately 2-fold). The expression assay of each chaperone and PHA synthase subunit (PhaRYB4 and PhaCYB4) indicated that the combination of the two chaperone systems (GroEL-GroES and TF) supported the folding of PhaRYB4 and PhaCYB4. These results suggest that the utilization of chaperone proteins is a valuable approach to enhance the formation of active PHA synthase and the productivity of 3HBO.

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“…Recently, it has been reported that PHA-producing E. coli secretes hydroxyalkanoate (HA) oligomers when an extremely high concentration (such as g/L) of diethylene glycol (DEG) was added as a CT agent to the culture medium (Utsunomia et al, 2017b, Utsunomia, et al, 2017c. The secretory production of HA oligomers has been demonstrated using Luria-Bertani (LB) medium supplemented with sugars as a carbon source, with DEG as the CT agent.…”

Section: Introductionmentioning

confidence: 99%

The influence of medium composition on the microbial secretory production of hydroxyalkanoate oligomers

Matsumoto

Sakurai

Nabasama

et al. 2021

J. Gen. Appl. Microbiol.

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With the aid of a chain transfer (CT) reaction, hydroxyalkanoate (HA) oligomers can be secreted by recombinant Escherichia coli carrying the gene encoding a lactate-polymerizing enzyme (PhaC1 Ps STQK) using Luria-Bertani (LB) medium supplemented with a carbon source and CT agent. In this study, HA oligomers were produced through microbial secretion using a mineral-based medium instead of LB medium, and the impact of medium composition on HA oligomer secretion was investigated. The focused targets were medium composition and NaCl concentration related to osmotic conditions. It was observed that 4.21 g/L HA oligomer was secreted by recombinant E. coli in LB medium, but the amount secreted in the mineral-based modified R (MR) medium was negligible. However, when the MR medium was supplemented with 5 g/L yeast extract, 3.75 g/L HA oligomer was secreted. This can be accounted for by the enhanced expression and activity of PhaC1 Ps STQK upon supplementation with growth-activated nutrients as supplementation with yeast extract also promoted cell growth and intracellular growth-associated polymer accumulation. Furthermore, upon adding 10 g/L NaCl to the yeast extract-supplemented MR medium, HA oligomer secretion increased to 6.86 g/L, implying that NaCl-induced osmotic pressure promotes HA oligomer secretion. These findings may facilitate the secretory

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Microbial secretion of lactate-enriched oligomers for efficient conversion into lactide: A biological shortcut to polylactide

Cited by 13 publications

References 26 publications

Microbial Secretion Platform for 3‐Hydroxybutyrate Oligomer and Its End‐Capped Forms Using Chain Transfer Reaction‐Mediated Polyhydroxyalkanoate Synthases

Microbial Secretion Platform for 3‐Hydroxybutyrate Oligomer and Its End‐Capped Forms Using Chain Transfer Reaction‐Mediated Polyhydroxyalkanoate Synthases

Enhanced Production of (R)-3-Hydroxybutyrate Oligomers by Coexpression of Molecular Chaperones in Recombinant Escherichia coli Harboring a Polyhydroxyalkanoate Synthase Derived from Bacillus cereus YB-4

The influence of medium composition on the microbial secretory production of hydroxyalkanoate oligomers

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