Production of polysaccharidases by a cellulase-pectinase hyperproducing mutant (Po16) of Penicillium occitanis S. Jain*, M. Parrichet, H. Durandt and G. Tiraby* *Laboratoire de Microbiologie et G#n#tique appliqudes du C. N. R. S., C. R. B. G. C., Universit# Paul Sabatier, ?Laboratoire CA YLA, Toulouse, France A new Penicillium strain characterized by high yields ofextracellular cellulase production, particularly fl-glucosidase, was isolated from soil. On the basis of growth characteristics, reverse coloration, and structure of Penicilli, this strain was assigned to a new species, named Penicillium occitanis. The organism was subjected to a strain-improvement program, and a mutant (Pol6) was selected after several rounds of mutagenesis with NTG, EMS, and UV. Selection was for strains that were resistant to catabolite repression and were enhanced in cellulase and pectinase production. The strain Po16 was also found to produce other polysaccharide-hydrolysing enzymes when grown on various inducing substrates. The production of polysaccharidases by Penicillium occitanis strain Pol6, under various fermentation conditions, and their preliminary separation by Fast Protein Liquid Chromatography is discussed. Keywords: Penicillium occitanis; Po16mutant; polysaccharidases;hydrolysis;mutagenesis;Fast Protein Liquid Chromatography(FPLC) Introduction Bioconversion of cellulosic wastes to useful products by enzymatic hydrolysis has been widely investigated for several years. ~-3High yields of cellulases are known to be produced by a few fungal genera, including Tri- choderma, Sporotrichum (Phanerochaete), Fusarium, Penicillium, and Talaromyces. The fungus Trichod- erma reesei makes an active cellulolytic complex com- prised of endo-l,4-fl-D-glucanases, exo-l,4-/3-D-cello- biohydrolases, and /3-D-glucosidase, which acts synergistically in the complete saccharification of cellu- lose to glucose. 1'4 Unfortunately, T. reesei produces only small amounts of/3-glucosidase,5 the enzyme re- quired for the final conversion of cellobiose to glucose. In contrast, P. funiculosurn produces high levels of extramycelial fl-glucosidase, in addition to cellobiohy- Addressreprintrequeststo Dr. Tiraby at the Laboratoire de Microbi- ologie et G6n6tique appliqu6es du C. N. R. S., C. R. B. G. C., Universit~ Paul Sabatier, 118 route de Narbonne, 31062Toulouse C6dex, France Received 22 May 1989; revised 14 August 1989 drolase and endo 1,4-fl-glucanases. 6 Furthermore, mu- tant strains of both P. funiculosum and P. pinophilum 7 have been developed for enhanced cellulase and /3- glucosidase production. However, complete hydroly- sis of raw materials also requires other polysacchari- dases such as xylanase, amylase, pectinase, and/3-1,3- glucanase. A new strain of Penicillum named P. occitanis was isolated from soil on the basis of its ability to produce large amounts of cellulases. This isolate is attractive because filtrates of cellulose-grown cultures have cot- ton-hydrolysing activities as high as those of T. reesei QM9414 and/3-glucosidase activities which are much higher) The isolate also produces a broad spectrum of polysaccharide-hydrolysing enzymes, which could be useful in the complete hydrolysis of complex raw sub- strates. The present study reports the isolation of a hyperse- creting mutant (Po16) ofPenicilliurn occitanis after mul- tiple-step mutagenesis and the production of polysac- charidases by this mutant when it is cultivated under various fermentation conditions. Anion-exchange chromatography was used for the preliminary separa- tion of the extracellular proteins. © 1990 Butterworth-Heineman Enzyme Microb. Technol., 1990, vol. 12, September 691