红豆中几丁质酶的纯化与表征-福州大学学报.PDFVIP

35 4 ( ) V o l 35 No 4 2007 8 Journa l of F uzhou U n iversity( N a tura l Sc ience) A ug 2007 : 1000 - 224 3( 2007) 04 - 0625 - 0 6 , , , , , (, 350002) : CM - Seph adex C- 50 Sephadex G - 75 (H PLC) Po ros H S- 20, SD S- PAGE 27 7 22 7 kD a, 14 57 U /m g, pH 5 4, 50 : ; ; ; : Q 814 1 : A Purification and characterization of a chitinase from the adzuki bean L IU X in - yi, XU E r- hua, CHEN Ji- w e,i L IU Y uan - gang, WANG Sh ao- yun, YE X iu - yun ( In stitute o f B iotechno logy, Fuzhou U n iversity, Fuzhou, Fu jian 350002, China) Abstract: A chitinase w ith antifungal act iv ity w as iso lated from adzu ki bean (Vigna angularis) seeds by a procedure conta in ing ( NH4 ) 2 SO4 precip itation, ion - exch ange chrom atog raphy on CM - Seph a dex C - 50, gel filtration on Seph adex G - 75, and h igh - p erform ance liqu id chrom atography ( H PLC ) on Po ros H S- 20 The protein exh ib ited a mo lecular m ass o f 27 7 kD a in redu ced cond ition w h ile 22 7 kD a in ox idized cond ition on SDS- PAGE, w h ich ind icates th at there are disu lfide bonds in side the m olecule T he specific activ ity of the chitinase w as estmi ated to be 14 57 U /m g The en zym e expressed its optmi um activ ity at pH 5 4 and a tem perature of 50 It exerted obv ious antifun gal act ion tow ards Fusarium oxysporum, Fusarium oxysporum f sp nelonis, and Pythium