Metabolic engineering of Bacillus subtilis for production of D-lactic acid

Awasthi, Deepika; Wang, Liang; Rhee, Mun S.; Wang, Qingzhao; Chauliac, Diane; Ingram, Lonnie O.; Shanmugam, Keelnatham T.

doi:10.1002/bit.26472

Metabolic engineering of Bacillus subtilis for production of D-lactic acid

Journal Article · Sat Oct 07 00:00:00 EDT 2017 · Biotechnology and Bioengineering

DOI:https://doi.org/10.1002/bit.26472· OSTI ID:1766311

Awasthi, Deepika ^[1]; Wang, Liang ^[2]; Rhee, Mun S. ^[2]; Wang, Qingzhao ^[2]; Chauliac, Diane ^[2]; Ingram, Lonnie O. ^[2]; ^[2]

Univ. of Florida, Gainesville, FL (United States); University of Florida
Univ. of Florida, Gainesville, FL (United States)

Poly lactic acid (PLA) based plastics is renewable, bio–based, and biodegradable. Although present day PLA is composed of mainly L–LA, an L– and D– LA copolymer is expected to improve the quality of PLA and expand its use. To increase the number of thermotolerant microbial biocatalysts that produce D–LA, a derivative of Bacillus subtilis strain 168 that grows at 50°C was metabolically engineered. Since B. subtilis lacks a gene encoding D–lactate dehydrogenase (ldhA), five heterologous ldhA genes (B. coagulans ldhA and gldA101, and ldhA from three Lactobacillus delbrueckii) were evaluated. Corresponding D–LDHs were purified and biochemically characterized. Among these, D–LDH from L. delbrueckii subspecies bulgaricus supported the highest D…LA titer (about 1M) and productivity (2 g h^–1 g cells^–1) at 37°C (B. subtilis strain DA12). The D–LA titer at 48°C was about 0.6 M at a yield of 0.99 (g D–LA g–1glucose consumed). Strain DA12 also fermented glucose at 48°C in mineral salts medium to lactate at a yield of 0.89 g g^–1 glucose and the D–lactate titer was 180 ± 4.5 mM. Furthermore, these results demonstrate the potential of B. subtilis as a platform organism for metabolic engineering for production of chemicals at 48°C that could minimize process cost.

View Accepted Manuscript (DOE)

Research Organization:: Univ. of Florida, Gainesville, FL (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE)

Grant/Contract Number:: PI0000031

OSTI ID:: 1766311

Alternate ID(s):: OSTI ID: 1408188

Journal Information:: Biotechnology and Bioengineering, Journal Name: Biotechnology and Bioengineering Journal Issue: 2 Vol. 115; ISSN 0006-3592

Publisher:: WileyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
Bacillus subtilis
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Fermentation
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Metabolic engineering of Bacillus subtilis for production of D-lactic acid

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