Preliminary Study On The Investigation, Classification And Producing Keratinase Of The Genus Chrysosporium

Fungi and their genetic resources and metabolic productions are invaluable biological resources. With the synergic development and mutual infiltration of different subjects, especially the molecular biology, more and more genetic resources and metabolic productions will be used. On the biotechnology market, fungi are widely used in many fields, such as medicine, foodstuff, agriculture, energy sources, fine chemicals, biological pesticides and environmental protection. Chrysosporium corda is a very important kind of fungal resource, which, according to the reports and the records, has more than 90 species, and the application value and the potential value had been reviewed in another paper. As the basic research of above all, the investigation and classification are getting more and more attention.Under the support of National Natural Science Foundation of China (30270011) and the instruction of my tutor, the resources of Chrysosporium corda were systematically studied, and the main results are as follows1. Acknowledgment of the globle valid species of Chrysosporium. The classification status and morphological characters of Chrysosporium were reviewed after systemically collecting and researching the materials about Chrysosporium all over the world; the results showed that there are 51 valid species in the genus Chrysosporium .2. the Access Database of Chrysoporium was set up according to its cultural characteristics and microscopic characteristics. The practices showed that the Access Database was convenient, quick and effective for identifying the strains.3. The investigation and classification to the resources of Chrysosporium in 13 different territories of China have been done. 36 strains of Chrysosporim from 438 soil samples were isolated and identified.Besides, there were 5 strains collected by the Institute of Fungal Resources of Gui Zhou University were also studied.The results described and identified mainly according to phenotype characteristics were as follows[1] A new record species of Chrysosporium in China:Chrysosporium tropicum J.W. Carmichael[2] 3 new specieses of ChrysosporiumChrysosporium pingwuense Z.Q.Liang, J.D.liang & Y.F.Han.sp.nov.Chrysosporium abaense Z.Q.Liang, J.D.liang & Y.F.Han.sp.nov.Chrysosporium yaoduense Z.Q.Liang, J.D.liang & Y.F.Han.sp.nov.[3] A new record species of Geomyces in ChinaGeomyces pannorum (Link) Sigler & Carmichael var. Pannorum4. The statistics showed that the distribution of Chrysosporium spp. was different between north and south in China, there was a diversity in the geography. The detection rate in north (10.3%) is higher than in south(5.9%), it implied the Chrysosporium spp. were cold-resistent, drought-resistent, stress-resistent and strongly adaptive, so Chrysosporium was a widely distributive group in geography.5. The research of keratinase-producing condition and the screening of keratinase-producing strains.According to Wawrzkiewicz's method, primary screening of keratinase-producing strains from some of the strains obtained by author has been done and inspected the results of primary screening using microscope. Keratinase activity of Chrysosporiwn pingwuense GZDXIFR-H-49-2 in ferment was also mensurated by UV method. Finally, the optimization of media and cultural condition were done by RSA method. The results is as follows[1] Obtaining Chrysosporiwn pingwuense GZDXIFR-H-49-2 with stronger ability producing keratinase by screening.[2] Under experiment condition, while the complements of C-sources, N-sources, S-sources and different mineral ions, the substrate with strongest promoter action were separately soluble starch, carbamide, hyposulphite and calcium chloride.[3] Obtaining quadratic polynomial by RSA method:Y1 = 5266.667 - 25 X₁ -162.5 X₂ - 212.5 X₃ -1058.333 X₁² -175 X₁ X₂ - 575 X₁ X₃ -1783.333 X₂² -1633.333 X₃²There was a Stationary point for Y1, while X₁= 0.010154, X₂= -0.046059, X₃= -0.066838, namely, feather powder 20.10g/L, carbamide0.9770 g/L, initial pH7.933 and another mineral ions(NaCl5g/L, K₂HPO₄ 6 g/L, KH₂PO₄ 1g/L), the prospective max value of keratinase activity was 5277.38 U/L. the prospective max value of keratinase activity was 6.6 times higher than the keratinase activity in ferment using the basic medium.