Purification, N-terminal amino acid sequencing and antifungal activity of chitinases from pepper stems treated with mercuric chloride
Purification, N-terminal amino acid sequencing and antifungal activity of chitinases from pepper stems treated with mercuric chloride
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氯化汞处理辣椒茎中几丁质酶的纯化、N 端氨基酸测序和抗真菌活性
DOI：
10.1006/pmpp.1996.0033
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Kook Hwang Byung
中科院分区：
文献类型：
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作者：
Jinchuan Young;Kook Hwang Byung
从辣椒(Capsicum annuum L.cv.)中分离纯化出不同亚型的几丁质酶。Hanbyul)茎用氯化汞处理。酸性异构体A1(69 kDa，pI5.0)、碱性异构体b1(32 kDa，pI9.0)和碱性异构体b2(22 kDa，pI9.1)经几丁质亲和层析纯化，然后从非变性的聚丙烯酰胺凝胶电泳胶(PAGE)中进行电洗脱。酸性异构体A1具有与甲壳素结合的特性，但没有抗真菌活性。基本异构体b1和b2在N-末端几丁质结合区含有高比例的半胱氨酸和甘氨酸，具有几丁质酶活性，并对炭疽菌、尖孢镰刀菌具有抗真菌活性。黄瓜、稻瘟病菌和绿色木霉的体外培养。此外，它们的抗真菌活性对丝状真菌表现出高度的特异性。几丁质酶b1和b2在其N端残基上与小麦、烟草、马铃薯、水稻和拟南芥的几丁质酶有很高的序列同源性。纯化的几丁质酶亚型均不能抑制缺少几丁质的辣椒疫霉卵菌的菌丝生长。相比之下，b1和b2处理能有效地抑制辣椒疫霉游动孢子萌发和芽管伸长。
Abstract Different isoforms of chitinases were purified from pepper ( Capsicum annuum L. cv. Hanbyul) stems treated with mercuric chloride. The acidic isoform a1 (69kDa, pI5.0), basic isoforms b1 (32kDa, pI9.0) and b2 (22kDa, pI9.1) were purified by chitin-affinity chromatography, with subsequent electroelution from nondenaturing polyacrylamide gel electrophoresis (PAGE) gels. The acidic isoform a1 has chitin-binding properties, but no antifungal activity. The basic isoforms b1 and b2 contain high ratios of cysteine and glycine at the N-terminal chitin-binding domain, exhibit chitinase activity, and show antifungal activities against Colletotrichum gloeosporioides, Fusarium oxysporum f.sp. cucumerinum, Magnaporthe grisea , and Trichoderma viride in vitro. Moreover, their antifungal activity shows a high degree of specificity to filamentous fungi. The chitinases b1 and b2 show a high sequence identity in their N-terminal residues with those from wheat, tobacco, potato, rice and Arabidopsis thaliana. None of the purified isoforms of chitinases inhibited hyphal growth of the Oomycete fungus which lacks chitin Phytophthora capsici . In contrast, zoospore germination and germ tube elongation of P. capsici were effectively inhibited by treatment with b1 and b2.