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Titolo:
Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production
Autore:
Chary, VK; de la Fuente, JL; Leitao, AL; Liras, P; Martin, JF;
Indirizzi:
Univ Leon, Fac Biol, Area Microbiol, E-24071 Leon, Spain Univ Leon Leon Spain E-24071 c Biol, Area Microbiol, E-24071 Leon, Spain
Titolo Testata:
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
fascicolo: 3, volume: 53, anno: 2000,
pagine: 282 - 288
SICI:
0175-7598(200003)53:3<282:OOTLGI>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
LYSINE EPSILON-AMINOTRANSFERASE; ALPHA-AMYLASE GENE; STREPTOMYCES-CLAVULIGERUS; CEPHALOSPORIN BIOSYNTHESIS; ANTIBIOTIC BIOSYNTHESIS; PURIFICATION; CLUSTER; EXPRESSION; GRISEUS; ENZYMES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
44
Recensione:
Indirizzi per estratti:
Indirizzo: Martin, JF Univ Leon, Fac Biol, Area Microbiol, E-24071 Leon, Spain Univ Leon Leon Spain E-24071 a Microbiol, E-24071 Leon, Spain
Citazione:
V.K. Chary et al., "Overexpression of the lat gene in Nocardia lactamdurans from strong heterologous promoters results in very high levels of lysine-6-aminotransferase and up to two-fold increase in cephamycin C production", APPL MICR B, 53(3), 2000, pp. 282-288

Abstract

The level of lysine-6-aminotransferase (encoded by the lat gene), an enzyme that commits lysine to the cephamycin biosynthesis pathway, is very low in wild type Nocardia lactamdurans . Two lat overexpression systems (pAMEXlat and pSAFlat) were constructed to express the promoterless fat gene of N. lactamdurans from the strong promoters amyP (of the alpha-amylase gene) andsafP (of the secretion activating factor gene) of Streptomyces griseus. Both constructions led to very high levels of lysine-6-aminotransferase (between 8- and 15-fold) in the cells. Expression of (at from the amy) promoter was optimal in glycerol-containing medium and was negatively regulated by glucose. The high levels of lysine-6-aminotransferase resulted in a 50 200% increase in cephamycin C production in the standard fermentation conditions. Onset of cephamycin C biosynthesis occurred at the same time in control and in lat-overexpressing strains, but the cephamycin production rate was clearly higher in transformants overexpressing the lat gene. Furthermore, HPLC analysis of cephamycin C in the culture broths revealed an early depletion of biosynthetic intermediates and an accumulation of cephamycin C when the lat gene was overexpressed. These results indicate that lysine-6-aminotransferase activity is limiting for cephamycin C biosynthesis under some culture conditions.

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Documento generato il 01/10/20 alle ore 00:07:56