Mutagenesis Of Protoplast Of Trichoderma Koningii And The Way That Produce Alcohol From Corn Straw

The material was stalk of maize and wheat bran, the starting bacterium was a kind of Trichoderma koningii in this experiment. The mutagenesis of protoplast of Trichoderma koningii was done by UV. The method of the mutagenesis of protoplast was established. The character of furguses after mutagenesis was determined. In addition, an differential medium which can conveniently segregate cellulose decomposing microorganisms was improved. The method which determine FP cellulose activity was improved. The simple way which produce alcohol from corn straw was established and the economy benefit analysis was done.The starting bacterium was a kind of Trichoderma koningii which was saved by our laboratory, the Trichoderma koningii was inoculated in the medium made of corn straw / wheat bran = 1 : 4 and foster 32h at 28℃.The FP cellulose activity of starting bacterium was 13.40+1.58ug/min.mL. The enzyme liquid was made by the enzyme dissolved cell wall. The Trichoderma koningii was operated by the liquid. After 3 hours, the protoplast of Trichoderma koningii was formed. The mutagenesis of protoplast of Trichoderma koningii was done by UV and fourty furguses were gained. After fostering of five era and primary isolation, seven furguses were gained. After further isolation, two furguses were gained and were named ZHYl and ZHY2. ZHY1 was inoculated in the medium made of corn straw / wheat bran = 1:4 and foster 32h at 28℃. The FP cellulose activity of ZHYl was 36.76+1.54ug/min.mL. ZHY2 was inoculated in the medium made of corn straw / wheat bran =1:4 and foster 32h at 28℃. The FP cellulose activity of ZHY2 was 33.40+2.80Mg/min.mL. After mulriple compare and analysis of difference prominence.The FP cellulose activity of ZHYl and ZHY2 prominent difference than the starting bacterium.The component of primary isolation medium included (NH4)2SO42g, MgSO4 0.5g, K2HPO4 1g, NaCl 0.5g, cellulose powder 2g, Congo red 0.4g, agar 22g, water1000mL, nature pH(the cellulose powder was operated by 0. lmol/I hydrochloric acid. After 24 hours, the cellulose powder was washed by water to the pH equal to 7. The agar was washed by water.After 24 hours, the agar was dryed.). The effect of medium is obviouser than the medium which was reported by literature. The cellulose activity of furguses was simply estimated by the size of the red colonies and the time which the red colonies appear.During determining of cellulose activity, the way distilled enzyme liquid was in shaking bed 1 hours at 30℃ and the revolution of shaking bed was 95 r/min. The distilling of the cellulase was more sufficient and the result of the cellulose activity was accurater by using this way.The way which made the Trichoderma koningii decompose the cellulose directly to produce alcohol was determined. This way is vary simple, and the way can produce alcohol from corn straw in laboratory.158g alcohol can be transformed by 1 kg corn straw and 4 kg wheat bran.