Stimulatory effects of chitinase on growth and immune defense of orange-spotted grouper (Epinephelus coioides).
Stimulatory effects of chitinase on growth and immune defense of orange-spotted grouper (Epinephelus coioides).
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几丁质酶对橙斑石斑鱼（Epinephelus coioides）生长和免疫防御的刺激作用。
DOI：
10.1016/j.fsi.2012.02.009
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发表时间：
2012-05
期刊：
影响因子：
--
通讯作者：
Li W
中科院分区：
文献类型：
--
作者：
Zhang Y;Feng S;Chen J;Qin C;Lin H;Li W
几丁质酶属于糖基水解酶家族18或家族19，将几丁质水解成寡糖。本研究将斜带石斑鱼几丁质酶1（Chitinase 1，Chitinase 1）的cDNA片段亚克隆到pPIC 3.5K载体中，并在毕赤酵母GS115中表达。结果表明，SDS-PAGE和Western blot均能检测到约53 kDa的条带。将重组石斑鱼几丁质酶1（rgChi 1）蛋白添加到含有虾壳几丁质的饲料中进行为期8周的饲养试验。在第2、4、6、8周分别测定饲料中rgChi 1含量为0、5、10、20 μg/g时的体重，在第6周观察到生长速度的差异，并一直持续到试验结束。在8周的饲养试验结束时，与对照组相比，添加10和20 μg/g饲料的鱼的增重百分比（PWG）、生长速率（GR）和特定生长速率（SGR）显著提高。添加10 μg/g或/和20 μg/g rgChi 1的鱼，其神经肽Y（NPY）、生长激素释放激素（GHRH）、生长激素（GH）、白细胞介素1 β（IL-1β）、环氧合酶2（考克斯-2）、超氧化物歧化酶（SOD）（Cu/Zn）和SOD（Mn）mRNA表达量均显著高于对照组。添加10和20 μg/g rgChi 1的鱼的溶菌酶（LZM）和总SOD活性显著高于对照组。20 μg/g组的谷草转氨酶（AST）/谷草转氨酶（GOT）活性较对照组降低。结果表明，利用毕赤酵母表达系统成功表达石斑鱼几丁质酶1，重组蛋白具有明显的促生长和免疫防御作用。石斑鱼几丁质酶1和几丁质酶2的mRNA表达和蛋白分泌在脾脏中的细菌脂多糖（LPS）的挑战显着刺激，强烈表明存在一个先天的途径，对含几丁质的生物体的局部防御。重组蛋白对大肠杆菌、金黄色葡萄球菌等病原菌有一定的抑制作用。
Chitinase, belonging to either family 18 or family 19 of the glycosylhydrolases, hydrolyze chitin into oligosaccharides. In the present study, the cDNA fragment encoding orange-spotted grouper (Epinephelus coioides) chitinase1 was subcloned into pPIC3.5K vector and expressed in Pichia pastoris GS115. The results showed that a band with the size of about 53 kDa could be detected by SDS-PAGE and Western blot. The recombinant protein of grouper chitinase1 (rgChi1) was added into the fish diet containing shrimp shell chitin for feeding experiment lasting 8 weeks. The weight of orange-spotted grouper, fed with diets containing rgChi1 at 0, 5, 10 and 20 μg/g was calculated on the 2nd, 4th, 6th and 8th weeks, and difference in growth rates was first observed in the 6th week of the feeding period and it kept until the end of the feeding experiment. At the end of 8 weeks feeding trial, the percent weight gain (PWG), growth rate (GR) and specific growth rate (SGR) of fish fed with 10 and 20 μg rgChi1/g feed were significantly higher compared to the control group. The neuropeptide Y (NPY), growth-hormone-releasing hormone (GHRH), growth-hormone (GH), interleukin-1beta (IL-1β), cyclooxygenase-2 (COX-2), superoxide dismutase (SOD) (Cu/Zn) and SOD (Mn) mRNA expression of fish fed with diet containing 10 μg/g or/and 20 μg/g rgChi1 were obviously higher than the control group. The lysozyme (LZM) and total SOD activity of fish fed with diet containing rgChi1 at 10 and 20 μg/g were significantly higher than that of the control. The aspartate aminotransferase (AST)/glutamic oxalacetic transaminases (GOT) activity in 20 μg/g group decreased compared to the control group. These results indicated that the grouper chitinase1 was successfully produced using the P. pastoris expression system and the recombinant protein had obvious effects on growth and immune defense. The mRNA expression and protein secretion of grouper chitinase1 and chitinase2 were significantly stimulated in spleen in response to bacterial lipopolysaccharide (LPS) challenge, strongly suggesting the existence of an innate pathway for local defense against chitin-containing organisms. Moreover, the pathogen such as Escherichia coli and Staphylococcus aureus could be inhibited by the recombinant protein of grouper chitinase1 to a certain extent.
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影响因子：
4.5
作者：
Gatesoupe, F. J.
通讯作者：
Gatesoupe, F. J.
DOI：
10.1016/s1096-4959(02)00046-5
发表时间：
2002-06
期刊：
Comparative biochemistry and physiology. Part B, Biochemistry & molecular biology
影响因子：
--
作者：
H. Oshima;R. Miyazaki;Y. Ohe;H. Hayashi;K. Kawamura;S. Kikuyama
通讯作者：
H. Oshima;R. Miyazaki;Y. Ohe;H. Hayashi;K. Kawamura;S. Kikuyama
影响因子：
4.8
作者：
RENKEMA, GH;BOOT, RG;AERTS, JMFG
通讯作者：
AERTS, JMFG
影响因子：
2.7
作者：
M. Mazumdar;B. Lal;A. Sakharkar;M. Deshmukh;P. Singru;N. Subhedar
通讯作者：
M. Mazumdar;B. Lal;A. Sakharkar;M. Deshmukh;P. Singru;N. Subhedar
DOI：
10.1007/978-3-0348-8757-1_11
发表时间：
1999
期刊：
--
影响因子：
--
作者：
Gooday Gw
通讯作者：
Gooday Gw