Improvement of<scp>L</scp>-Lactate Production by<i>CYB2</i>Gene Disruption in a Recombinant<i>Saccharomyces cerevisiae</i>Strain under Low pH Condition

Ookubo, Aki; Hirasawa, Takashi; Yoshikawa, Katsunori; Nagahisa, Keisuke; Furusawa, Chikara; Shimizu, Hiroshi

doi:10.1271/bbb.80493

Cited by 22 publications

(9 citation statements)

References 14 publications

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“…Gene disruption using the disruption cassette was performed based on the method by Güldener et al (1996). The HIS3 deletion cassette with homologous regions for each target gene was amplified by PCR from pUG6–HIS3 (Ookubo et al , 2008) using Z ‐ Taq polymerase (Takara Bio Inc., Shiga, Japan) and primer Nos 104 and 105 for ATH1 , 106 and 107 for NTH1 , and 108 and 109 for NTH2 (Table 1). After that, each amplified cassette was introduced into the FY834 strain by the lithium acetate method (Gietz and Woods, 2002) and the transformants were selected on SC − His plates.…”

Section: Methodsmentioning

confidence: 99%

Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae

Mahmud

Nagahisa

Hirasawa

et al. 2009

Yeast

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To examine the effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae, we constructed deletion strains of all combinations of the trehalase genes ATH1, NTH1 and NTH2 and examined their growth behaviour and intracellular trehalose accumulation under non-stress and saline-stress conditions. Saline stress was induced in yeast cells by NaCl addition at the exponential growth phase. All deletion strains showed similar specific growth rates and trehalose accumulation to their parent strain under non-stress conditions. However, under the saline stress condition, one single deletion strain, nth1 , two double deletion strains, nth1 ath1 and nth1 nth2 , and the triple deletion strain nth1 nth2 ath1 , all of which carry the nth1 deletion, showed increased trehalose accumulation as compared to the parent and other deletion strains. In particular, our statistical analysis revealed that the triple deletion strain showed a higher growth rate under the saline stress condition than the parent strain. Moreover, some deletion strains showed further trehalose accumulation under non-stress conditions by overexpression of the TPS1 or TPS2 genes encoding the enzymes related to trehalose biosynthesis at the mid-exponential phase. Such increased trehalose accumulation prior to NaCl addition could improve the growth of these strains under saline stress. Our results indicate that high trehalose accumulation prior to NaCl addition, rather than after NaCl addition, is necessary to achieve high growth activity under stress conditions.

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Section: Methodsmentioning

confidence: 99%

Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae

Mahmud

Nagahisa

Hirasawa

et al. 2009

Yeast

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“…L-lactate-cytochrome c oxidoreductase (or cytochrome b2, CYB2) is a soluble protein from the inter-membrane space of mitochondria which catalyses the oxidation of L-lactate into pyruvate and the transfer of electrons to cytochrome c. it has been shown, that CYB2 is up regulated by lactate, but repressed by glucose (Ramil et al 2000). Ookubo et al set out to analyse lactic acid production in a CYB2 deletion strain of S. cerevisiae (Ookubo et al 2008). interestingly, they showed that CYB2 deletion has no significant impact on lactate yield at high pH.…”

Section: Specific Metabolic Traits In the Limelight For Lactic Acid Pmentioning

confidence: 99%

16 years research on lactic acid production with yeast – ready for the market?

Sauer

Porro

Mattanovich

et al. 2010

Biotechnology and Genetic Engineering Reviews

110

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The use of plastic produced from non-renewable resources constitutes a major environmental problem of the modern society. Polylactide polymers (PLA) have recently gained enormous attention as one possible substitution of petroleum derived polymers. A prerequisite for high quality PLA production is the provision of optically pure lactic acid, which cannot be obtained by chemical synthesis in an economical way. Microbial fermentation is therefore the commercial option to obtain lactic acid as monomer for PLA production. However, one major economic hurdle for commercial lactic acid production as basis for PLA is the costly separation procedure, which is needed to recover and purify the product from the fermentation broth. Yeasts, such as Saccharomyces cerevisiae (bakers yeast) offer themselves as production organisms because they can tolerate low pH and grow on mineral media what eases the purification of the acid. However, naturally yeasts do not produce lactic acid. By metabolic engineering, ethanol was exchanged with lactic acid as end product of fermentation. A vast amount of effort has been invested into the development of yeasts for lactic acid production since the first paper on this topic by Dequin and Barre appeared 1994. Now yeasts are very close to industrial exploitation - here we summarize the developments in this field.

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“…S . cerevisiae utilizes L-lactate via a mitochondrial L(+)-lactate:cytochrome c oxidoreductase (encoded by CYB2 ) and disruption of CYB2 improved the lactate production [ 48 ]. Lactate yield marginally improved in A .…”

Section: Resultsmentioning

confidence: 99%

Expression of Lactate Dehydrogenase in Aspergillus niger for L-Lactic Acid Production

Dave

Punekar

2015

PLoS ONE

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Different engineered organisms have been used to produce L-lactate. Poor yields of lactate at low pH and expensive downstream processing remain as bottlenecks. Aspergillus niger is a prolific citrate producer and a remarkably acid tolerant fungus. Neither a functional lactate dehydrogenase (LDH) from nor lactate production by A. niger is reported. Its genome was also investigated for the presence of a functional ldh. The endogenous A. niger citrate synthase promoter relevant to A. niger acidogenic metabolism was employed to drive constitutive expression of mouse lactate dehydrogenase (mldhA). An appraisal of different branches of the A. niger pyruvate node guided the choice of mldhA for heterologous expression. A high copy number transformant C12 strain, displaying highest LDH specific activity, was analyzed under different growth conditions. The C12 strain produced 7.7 g/l of extracellular L-lactate from 60 g/l of glucose, in non-neutralizing minimal media. Significantly, lactate and citrate accumulated under two different growth conditions. Already an established acidogenic platform, A. niger now promises to be a valuable host for lactate production.

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Improvement ofL-Lactate Production byCYB2Gene Disruption in a RecombinantSaccharomyces cerevisiaeStrain under Low pH Condition

Cited by 22 publications

References 14 publications

Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae

Effect of trehalose accumulation on response to saline stress in Saccharomyces cerevisiae

16 years research on lactic acid production with yeast – ready for the market?

Expression of Lactate Dehydrogenase in Aspergillus niger for L-Lactic Acid Production

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