Detection and production of extracellular collagenolytic enzyme from Zygosaccharomyces rouxii.

Abstract

Microbial collagenases are proteinases that are capable of hydrolyzing native collagen in vitro under physiological conditions ( 7) . Production of collagenase or collagenase-like proteases from various bacteria have been reported by many workers. Among the collagenase-producing bacteria, Clostridium histolyticum is the most studied microorganism and crude or purified collagenase from this bacteria are now available commercially. There have been works published on collagenase from the mold Trichophyton shoenleinii (13) and Aspergillus oryzae (12). However, reports on the presence of collagenolytic activity in yeasts are very rare. In the book, "The Yeasts: A Taxonomic Study," it is stated that with the exception of Cryptococcus species, few yeasts have the ability to liquefy gelatin which is a denatured form of collagen (5). Through our identification of sugartolerant yeasts isolated from high-sugar foods, we found a few strains of Zygosaccharomyces rouxii that possessed the activity of gelatin liquefaction. Although the enzyme active against gelatin may not be active against collagen, as pointed out by Merkel et al. (9), we were prompted by our results and regarded that it would be beneficial to investigate the presence of collagenolytic enzyme from a different source. This paper reports the isolation of crude enzyme from a gelatin-liquefaction strain of yeast and determination of its activity against various substrates including insoluble collagen. We dealt also with the cultural conditions that affected the production of the enzyme. Comparison was made between the yeast enzyme and Cl. histolyticum collagenase.Sugar-tolerant yeasts including 19 strains of Z. rouxii and one strain of Torulaspora delbrueckii, isolated from high-sugar foods were used for screening

Collagenolytic enzymes produced by fungi: a systematic review

“…Some studies involving bacteria indicate that adding gelatin or casein in the medium increases the collagenase yield. However, the work of Ok and Hashinaga 48 with Z. rouxii yeast, observed that adding gelatin in YPG medium was not essential for the production of collagenase. Lima et al 24 reported the use of a inexpensive culture medium for P. aurantiogriseum collagenase production, using soy flour as main substrate, and the same medium was used by authors Lima et al, 58 reaching one of the best collagenolytic activity values found during this review (Table 3).…”

“…For the best results regarding collagenolytic activity, Lima et al 58 and Mahmoud et al, 53 the optimum pH of the enzyme was not evaluated. Ok and Hashinaga 48 evaluated the optimal pH (8.2) of the enzyme produced by Z. rouxii yeast. Lima et al 24 found that pH of 9.0 was the best for collagenolytic enzyme produced by P. aurantiogriseum .…”

Production and Characterization of Collagenase by Penicillium sp. UCP 1286 Isolated From Caatinga Soil

“…This value is much higher than other published results. Baehaki et al [35] achieved a specific activity of 0.546 U/mg, with Bacillus licheniformis; Jain and Jain [1] [37], 70.4 U/mg with Zygosaccharomyces rouxii, all of them with more than 15 times lower activity than the produced by Penicillium sp. UCP 1286.…”

“…Lima et al [9] showed that the enzyme produced by P. aurantiogriseum was very active at the pH range 8 to 10, and the highest activity occurred at pH 9.0, as in the present work. Ok and Hashinaga [37] related that optimum pH to collagenase produced by Zygosaccharomyces rouxii was 8.2. Only the enzyme produced by Rhizoctonia solani was produced under acid pH (5.0) [14].…”

“…Hamdy [14] related specificity of collagenase produced by R. solani to gelatin and collagen (33.23 U/mg and 120 U/mg, respectively). Ok and Hashinaga [37] tested collagenase activity using soluble (27.1 U/mL) and insoluble collagen (5.6 U/mL), besides synthetic peptides as Cbz-GPLGP (21.1 U/mL) and FALGPA (0.41 U/mL). The collagenase produced by B. licheniformis exhibited the highest activity on casein, being also able to hydrolyze collagen, gelatin and fibrin [35].…”