폐글리세롤을 탄소원으로 hemA와 otsBA를 공동 발현하는 재조합 대장균 배양을 통한 아미노레블린산 생산
Production of Aminolevulinic Acid by Recombinant Escherichia coli Co-expressing hemA and otsBA Using Crude Glycerol as Carbon Source
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
This study aimed to develop a microbial process for producing aminolevulinic acid (ALA) using crude glycerol. In the culture of ALA-producing cells (Escherichia coli/pH-hemA) in a medium containing crude glycerol, the cell density and production were 1.8-fold and 1.2-fold lower than those obtained from pure glycerol, respectively. However, the cell growth and production were improved by supplementing the medium with trehalose (30 or 100 g/l). Engineered cells (E. coli/pH-hemA/pS-otsBA) were constructed to express otsBA and their culture performance was compared with that of control cells (E. coli/pH-hemA/ pSTV28). The effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and the time of induction were examined to improve the cell growth and ALA production in engineered cells cultured using crude glycerol. When 0.6 mM of IPTG was added at the beginning of the exponential growth phase, the ALA produced by cells was 2,121 mg/l, which was comparable to that from pure glycerol. The results demonstrate that otsBA expression endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.
Abstract
This study aimed to develop a microbial process for producing aminolevulinic acid (ALA) using crude glycerol. In the culture of ALA-producing cells (Escherichia coli/pH-hemA) in a medium containing crude glycerol, the cell density and production were 1.8-fold and 1.2-fold lower than those obtained from pure glycerol, respectively. However, the cell growth and production were improved by supplementing the medium with trehalose (30 or 100 g/l). Engineered cells (E. coli/pH-hemA/pS-otsBA) were constructed to express otsBA and their culture performance was compared with that of control cells (E. coli/pH-hemA/ pSTV28). The effects of isopropyl β-D-1-thiogalactopyranoside (IPTG) concentration and the time of induction were examined to improve the cell growth and ALA production in engineered cells cultured using crude glycerol. When 0.6 mM of IPTG was added at the beginning of the exponential growth phase, the ALA produced by cells was 2,121 mg/l, which was comparable to that from pure glycerol. The results demonstrate that otsBA expression endowed cells with the capacity to tolerate the toxicity of crude glycerol for direct use.
참고문헌 (39)
- Alvarez-Peral FJ, "Protective role of trehalose during severe oxidative stress caused by hydrogen peroxide and the adaptive oxidative stress response in Candida albicans", Microbiol, 148, pp.2599~2606 (2002)
- Benaroudj N, "Trehalose accumulation during cellular stress protects cells and cellular proteins from damage by oxygen radicals", J. Biol. Chem, 276, pp.24261~24267 (2001)
- Cabiscol E, "Oxidative stress in bacteria and protein damage by reactive oxygen species", Int. Microbiol, 3, pp.3~8 (2000)
- Chatzifragkou A, "Effect of biodiesel-derived waste glycerol impurities on biomass and 1,3-propanediol production of Clostridium butyricum VPI 1718", Biotechnol. Bioeng, 107, pp.76~84 (2010)
- Choi, Han-Pil, "Extracellular 5-Aminolevulinic Acid Production by Escherichia coli Containingthe Rhodopseudomonas palustris KUGB306 hemA Gene", Journal of Microbiology and Biotechnology, 18(6), pp.1136~1140 (2008), 한국미생물·생명공학회
- Dobson R, "Microbial utilization of crude glycerol for the production of value-added products", J. Ind. Microbiol. Biotechnol, 39, pp.217~226 (2012)
- Donovan RS, "Review: optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter", J. Ind. Microbiol, 16, pp.145~154 (1996)
- Fay JP, "The inhibitory action of fatty acids on the growth of Escherichia coli", J. Gen. Microbiol, 91, pp.233~240 (1975)
- Fotinos N, "5-aminolevulinic acid derivatives in photomedicine; characteristics, application and perspectives", Photochem. Photobiol, 82, pp.994~1015 (2006)
- Havasi V, "Inhibitory effects of hypertonic saline on P. aeruginosa motility", J. Cyst. Fibros, 7, pp.267~269 (2008)
- Herdeiro RS, "Trehalose protects Saccharomyces cerevisiae from lipid peroxidation during oxidative stress", Biochim. Biophys. Acta, 1760, pp.340~346 (2006)
- Imlay JA, "Cellular defenses against superoxide and hydrogen peroxide", Annu. Rev. Biochem, 77, pp.755~776 (2008)
- Ingram LO, "Adaptation of membrane lipids to alcohols", J. Bacteriol, 125, pp.670~678 (1976)
- Iturriaga G, "Trehalose metabolism:from osmoprotection to signaling", Int. J. Mol. Sci, 10, pp.3793~3810 (2009)
- KANG, DAE-KYUNG, "Cloning and Expression of the Rhodobacter capsulatus hemA Gene in E. colifor the Production of 5-Aminolevulinic Acid", Journal of Microbiology and Biotechnology, 14(6), pp.1327~1332 (2004), 한국미생물·생명공학회
- Kempf B, "Uptake and synthesis of compatible solutes as microbial stress responses to high-osmolality environments", Arch. Microbiol, 170, pp.319~330 (1998)
- Li C, "Escherichia coli shows two types of behavioral responses to osmotic upshift", J. Bacteriol, 175, pp.2564~2567 (1993)
- Li H, "Enhanced production of trehalose in Escherichia coli by homologous expression of otsBA in the presence of the trehalase inhibitor, validamycin A, at high osmolarity", J. Biosci. Bioeng, 113, pp.224~232 (2012)
- Liu X, "Bioconversion of crude glycerol feedstocks into ethanol by Pachysolen tannophilus", Biores. Technol, 104, pp.579~586 (2012)
- Luo Y, "Trehalose: protector of antioxidant enzymes or reactive oxygen species scavenger under heat stress?", Environ. Exp. Bot, 63, pp.378~384 (2008)
- Maia MRG, "Toxicity of unsaturated fatty acids to the biohydrogenating ruminal bacterium, Butyrivibrio fibrisolvens", BMC. Microbiol, 10, pp.52~ (2010)
- Nery DCM, "The role of trehalose and its transporter in protection against reactive oxygen species", Biochim. Biophys. Acta, 1780, pp.1408~1411 (2008)
- Nguyen ADQ, "Improved tolerance of recombinant Escherichia coli to the toxicity of crude glycerol by overexpressing trehalose biosynthetic genes (otsBA) for the production of β-carotene", Biores. Technol, 143, pp.531~537 (2013)
- Nishikawa S, "Rhodobacter sphaeroides mutants which accumulate 5-aminolevulinic acid under aerobic and dark conditions", J. Biosci. Bioeng, 87, pp.798~804 (1999)
- Oku K, "NMR and quantum chemical study on the OH-π and CH-O interactions between trehalose and unsaturated fatty acids: implication for the mechanism of antioxidant function of trehalose", J. Am. Chem. Soc, 125, pp.12739~12748 (2003)
- Papaneophytou CP, "Statistical approach to maximize recombinant protein expression in Escherichia coli: a general review", Protein Expr. Purif, 94, pp.22~32 (2014)
- Purvis JE, "Enhanced trehalose production improves growth of Escherichia coli under osmotic stress", Appl. Environ. Microbial, 71, pp.3761~3769 (2005)
- Rodriguez LC, "Damage of Escherichia coli cells by t-butylhydroperoxide involves the respiratory chain but is independent of the presence of oxygen", Biochim. Biophys. Acta, 1015, pp.510~516 (1990)
- Ruhal R, "Trend in bacterial trehalose metabolism and significant nodes of metabolic pathway in the direction of trehalose accumulation", Microbiol. Biotechnol, 6, pp.493~502 (2013)
- Samul D, "Impurities of crude glycerol and their effect on metabolite production", Ann. Microbiol, 64, pp.891~898 (2014)
- Sasikala C, "5-aminolevulinic acid: a potential herbicide", insecticide from microorganisms, 1994, pp.1994, 451~459
- Shah P, "Aerobic utilization of crude glycerol by recombinant Escherichia coli for simultaneous production of poly 3-hydroxybutyrate and bioethanol", J. Biosci. Bioeng, 117, pp.343~350 (2014)
- Sivashanmugam A, "Practical protocols for production of very high yields of recombinant proteins using Escherichia coli", Protein Sci, 18, pp.936~948 (2009)
- Sugamata Y, "Improved secretory production of recombinant proteins by random mutagenesis of hlyB, an alphahemolysin transporter from Escherichia coli", Appl. Environ. Microbiol, 71, pp.656~662 (2005)
- Venkataramanan KP, "Impact of impurities in biodieselderived crude glycerol on the fermentation by Clostridium pasteurianum ATCC 6013", Appl. Microbiol. Biotechnol, 93, pp.1325~1335 (2012)
- Xie L, "Optimization of recombinant aminolevulinate synthase production in Escherichia coli using factorial design", Appl. Microbiol. Biotechnol, 63, pp.267~273 (2003)
- 양동수, "Rhodobacter capsulatus hemA 유전자 발현 대장균에 의한 5-Aminolevulinic Acid 생산의 최적화", 한국미생물·생명공학회지, 37(2), pp.153~159 (2009), 한국미생물·생명공학회
- Yang F, "Value-added uses for glycerol-a byproduct of biodiesel production", Biotechnol. Biofuels, 5, pp.13~ (2012)
- Zhang J, "Optimization of the heme biosynthesis pathway for the production of 5-aminolevulinic acid in Escherichia coli", Sci. Rep, 5, pp.8584~ (2015)