RRC ID |
49575
|
著者 |
Ozaki A, Konishi R, Otomo C, Kishida M, Takayama S, Matsumoto T, Tanaka T, Kondo A.
|
タイトル |
Metabolic engineering of Schizosaccharomyces pombe via CRISPR-Cas9 genome editing for lactic acid production from glucose and cellobiose.
|
ジャーナル |
Metab Eng Commun
|
Abstract |
Modification of the Schizosaccharomyces pombe genome is often laborious, time consuming due to the lower efficiency of homologous recombination. Here, we constructed metabolically engineered S. pombe strains using a CRISPR-Cas9 system and also demonstrated D-lactic acid (D-LA) production from glucose and cellobiose. Genes encoding two separate pyruvate decarboxylases (PDCs), an L-lactic acid dehydrogenase (L-LDH), and a minor alcohol dehydrogenase (SPBC337.11) were disrupted, thereby attenuating ethanol production. To increase the cellular supply of acetyl-CoA, an important metabolite for growth, we introduced genes encoding bacterial acetylating acetaldehyde dehydrogenase enzymes (Escherichia coli MhpF and EutE). D-LA production by the resulting strain was achieved by expressing a Lactobacillus plantarum gene encoding D-lactate dehydrogenase. The engineered strain efficiently consumed glucose and produced D-LA at 25.2 g/L from 35.5 g/L of consumed glucose with a yield of 0.71 g D-LA / g glucose. We further modified this strain by expressing beta-glucosidase by cell surface display; the resulting strain produced D-LA at 24.4 g/L from 30 g/L of cellobiose in minimal medium, with a yield of 0.68 g D-LA / g glucose. To our knowledge, this study represents the first report of a S. pombe strain that was metabolically engineered using a CRISPR-Cas9 system, and demonstrates the possibility of engineering S. pombe for the production of value-added chemicals.
|
巻・号 |
5
|
ページ |
60-67
|
公開日 |
2017-12-1
|
DOI |
10.1016/j.meteno.2017.08.002
|
PII |
S2214-0301(17)30014-7
|
PMID |
29188185
|
PMC |
PMC5699526
|
リソース情報 |
遺伝子材料 |
pDUAL-FFH61 (RDB06538)
pDUAL-HFG31 (RDB06540). |