Regulation of cellulases and xylanases from a derepressed mutant of Cellulomonas flavigena growing on sugar-cane bagasse in continuous culture.
When the wild type Cellulomonas flavigena was grown on glycerol, xylose or cellobiose, it produced basal levels of carboxymethyl-cellulase (CMCase), filter-paperase (FPase) and xylanase activities. By comparison, a catabolic derepressed mutant strain of the same organism produced markedly higher levels of these enzymes when grown on the same carbon sources. Sugar-cane bagasse induced both the wild type and the mutant strain to produce three- to eight-time higher levels of FPase and xylanase than was observed with xylose or cellobiose. Continuous culture was used to determine the minimal cellobiose or glucose concentrations that repress the enzyme synthesis in both strains. 2.5 g l(-1) glucose repressed FPase and xylanases from wild type, while 1.6 times more glucose was needed to repress the same activities in the PN-120 strain. In the same way, twofold more cellobiose was needed to reduce by 75% the CMCase and xylanase activities in the mutant compared to the wild type. The FPase in the presence of 4 g l(-1) cellobiose did not change in the same strain. Therefore, its derepressed and feedback resistant characters of PN-120 mutant are evident. On the other hand, isoelectrofocused crude extracts of mutant and wild strains induced by sugar-cane bagasse, did not show differences in protein patterns, however, the Schiffs staining was more intense in the PN-120 than in the wild strain. These results point out that the mutational treatment did not apparently change the extracellular proteins from mutant PN-120 and this could affect their regulation sites, since derepressed and feed-back resistant enzymes may be produced.
Ponce-Noyola
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T
Department of Biotechnology and Bioengineering, CINVESTAV-IPN, Col Zacatenco, DF, Mexico. tponce@mail.cinvestav.mx
de la Torre
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M
D000192:Actinomycetales / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D002480:Cellulase / Q000096:biosynthesis / Q000378:metabolism*; D002482:Cellulose; D004794:Enzyme Repression; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000378:metabolism; D007700:Kinetics; D016296:Mutagenesis; D010209:Paper; D013379:Substrate Specificity; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000096:biosynthesis / Q000378:metabolism*
D002482:Cellulose; C027433:bagasse; D006026:Glycoside Hydrolases; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase; D002480:Cellulase; C019826:carboxymethylcellulase