PURPOSE: To obtain a novel DNA coding an endo type inulinase hydrolyzing inuline to give inulooligosaccharide, and capable of producing the inulooligosaccharide in a high yield by inserting the novel DNA into a proper vector, transforming a microorganism, etc., with the vector and subsequently culturing the microorganism, etc.
CONSTITUTION: Penicillum purpurogenum var. rubrischerotium (FERM BP-3162) is cultured in a medium containing inuline, and all RNAs are extracted from the cultured cells by a guanicline chloride/phenol method. A mRNA is separated from all the RNAs with an oligo (dT) column, and a cDNA is synthesized with the mRNA as a template. The library of the cDNA is made by a conventional method, and subsequently screened with a probe prepared by synthesizing a part of a DNA coding endo-type inulinase. A positive clone is selected, and the DNA is recovered and subsequently treated with a proper restriction enzyme to obtain the objective novel DNA coding the endo type inulinase capable of hydrolyzing the inuline to produce inulooligosaccharide.
SHIOMI TOKUO
NAKAMURA TAKESHI
FUKUOKA AKIO
YOSHIMI FUMINOBU
