降解PHBV菌株的选育及其酶学性质的研究-微生物学专业论文.docxVIP

AbstractPolyf Abstract Polyf B·Hydroxybutyrate*CO-13-Hydroxyvalerate)(PHBV)are biodegradable polysters that are synthesized and accumulated intraeellularly during unbalanced growth by some bacteria．It has attracted much commercial and academic interest aS a kind of new biodegradable material． The ability to degrade PHBV is widely distributed among bacteria and fungi，and depends On the secretion of specific extracellulal PHBV depolymerase which can hydrolyse the polymer to water-soluble products．The aim ofthis work is协select the mutant with the treatment ofUV which call degrade PHBV efficiently,to purify PHBV depolymerase and to study the fundamental characteristics of PHBV depolymerase．The main results obtained from this work ale∞follows： ．The pinicillium sp DS9702一D10,a strain of degrading糟BV was mutagenized by UV treatment．Through screening a lot of mutants with the method of transparent zones and culture filtrate，best five ones vcere obtained弼魄撼gh—yield ofstable PHBV depolymerase， named as 05，06，11，19 and H2．骶忙enzyme activity ofthe06and H2 were 5．6 and S．38 times ofthe original OIle。 ≥．The enzyme activity and the protein of the five muntantsⅥ㈣surveyed with their crude enzyme，the highest ellzylD_e yielding ofthem w雠found in 72h of 11，19；96h ofI-12 and 120hof05。06．Theactivityofll、19、H2、05、06were2．75；1．55；5．38．4．37：5．6times ofthe original one。 3．Comparative study among the crude extracts of 06、1-12 mutants and the original one。The optimuln temperature of 06 was 20℃higher and H2 Was JO℃higher than the original one， the range of temperature stability of 06 was from 40C to 60。C and that of H2 was from 30 ℃to 60℃。but the original one was from 30℃to 50℃．The optimnm pH of06 Was 7．38 and that ofH2 Was 6．98 and 8．04．It Was different from the original one which was 7．0，Moreover, the range ofpH stabifity of06 and 1-12 was also much better． 4．The extracellular PHBV depolymerase Was selected and purified from 06 and H2 mutants by using filtration、C2H50H precipitation、gel filt