Metabolic engineering of Bacillus subtilis for production of D-lactic acid
Abstract
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.
- Publication Date:
- Research Org.:
- Univ. of Florida, Gainesville, FL (United States)
- Sponsoring Org.:
- USDOE Office of Energy Efficiency and Renewable Energy (EERE)
- OSTI Identifier:
- 1766311
- Alternate Identifier(s):
- OSTI ID: 1408188
- Grant/Contract Number:
- PI0000031
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology and Bioengineering
- Additional Journal Information:
- Journal Volume: 115; Journal Issue: 2; Journal ID: ISSN 0006-3592
- Publisher:
- Wiley
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; D-lactic acid; Bacillus subtilis; Lactobacillus ldhA; D-LDH activity; Fermentation
Citation Formats
Awasthi, Deepika, Wang, Liang, Rhee, Mun S., Wang, Qingzhao, Chauliac, Diane, Ingram, Lonnie O., and Shanmugam, Keelnatham T. Metabolic engineering of Bacillus subtilis for production of D-lactic acid. United States: N. p., 2017.
Web. doi:10.1002/bit.26472.
Awasthi, Deepika, Wang, Liang, Rhee, Mun S., Wang, Qingzhao, Chauliac, Diane, Ingram, Lonnie O., & Shanmugam, Keelnatham T. Metabolic engineering of Bacillus subtilis for production of D-lactic acid. United States. https://doi.org/10.1002/bit.26472
Awasthi, Deepika, Wang, Liang, Rhee, Mun S., Wang, Qingzhao, Chauliac, Diane, Ingram, Lonnie O., and Shanmugam, Keelnatham T. Sat .
"Metabolic engineering of Bacillus subtilis for production of D-lactic acid". United States. https://doi.org/10.1002/bit.26472. https://www.osti.gov/servlets/purl/1766311.
@article{osti_1766311,
title = {Metabolic engineering of Bacillus subtilis for production of D-lactic acid},
author = {Awasthi, Deepika and Wang, Liang and Rhee, Mun S. and Wang, Qingzhao and Chauliac, Diane and Ingram, Lonnie O. and Shanmugam, Keelnatham T.},
abstractNote = {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.},
doi = {10.1002/bit.26472},
journal = {Biotechnology and Bioengineering},
number = 2,
volume = 115,
place = {United States},
year = {Sat Oct 07 00:00:00 EDT 2017},
month = {Sat Oct 07 00:00:00 EDT 2017}
}
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