摘要: |
为获得高产纤维降解酶瘤胃厌氧真菌,从放牧饲养条件下随机选出10头屠宰后的青海大通牦牛瘤胃中采集食糜样本,通过亨盖特滚管技术筛选得到29株厌氧真菌菌株。以小麦秸秆为底物,测定这些菌株第6天培养液中乙酰酯酶、阿魏酸酯酶、木聚糖酶、羧甲基纤维素酶和微晶纤维素酶的活性。29株真菌之间5种酶活性差异显著,阿魏酸酯酶活性最高的菌株比活性最低的菌株活性高出7.9倍,差异最显著;而羧甲基纤维素仅高出2.5倍,差异最不显著。对各菌株5种酶活性进行相关分析,乙酰酯酶、阿魏酸酯酶、木聚糖酶和羧甲基纤维素酶相互之间均存在显著的正相关性(r>0.67,P<0.000 1),而且它们之间的相关性高于其与微晶纤维素间的相关性(P>0.05)。从中筛选得到1株高产纤维降解酶厌氧真菌YAK11,该菌株第6天培养液中5种酶活性分别达到165、10、1138、83和16 mU。通过18S rDNA基因序列分析该株菌,初步确定为Neocallimastix frontalis。 |
关键词: 牦牛 纤维降解酶 亨盖特滚管技术 18S rDNA 瘤胃 厌氧真菌 |
DOI:10.11841/j.issn.1007-4333.2010.03.011 |
投稿时间:2009-11-18 |
基金项目:国家自然科学基金资助项目(30400315); 农业部公益性行业科研专项(200803033B050401); 北京市大学生科学研究与创业行动计划第一作者:曹阳春,博士研究生,E-mail:caoyangchun@126.com 通讯作者:杨红建,副教授,博士生导师,主要从事反刍动物营养与饲料资源开发研究,E-mail:yang_hongjian@sina.com |
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Screening and identification of high activity cellulolytic anaerobicfungi from rumen of yak |
CAO Yang-chun1, YANG Hong-jian1,2, SHEN Bo-tong1
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(1.College of Biological Sciences,China Agricultural University,Beijing 100193,China;2.College of Animal Science and Technology,China Agricultural University,Beijing 100193,China) |
Abstract: |
The purpose of this study was to obtain superior cellulolytic anaerobic rumen fungi.Chyme samples were collected from rumen of ten slaughtered Datong yak, which were fed hay only in Qinghai.29 anaerobic fungi strains were isolated by the method of Hungate rolling tube technique.Activities of acetyl esterase, ferulic acid esterase,xylanase,carboxymethyl cellulose and avicelase at the sixth day of 29 fungi cultures were measured with wheat straw as substrate.The activities of five enzymes of the 29 fungi were significantly different,where ferulic acid esterase had substantially higher activity,which was 7.9 times higher than the enzyme with lowest activity.Activities of the four enzymes,acetyl esterase,ferulic acid esterase,xylanase and carboxymethyl cellulose,are highly correlated with each other(r>0.67,P<0.0001),while there was no such correlations observed with avicelase (P>0.05).A high activity anaerobic fungi,YAK11,was obtained, achieved the activities of 165,10,1138,83 and 16 mU for the five enzymes,respectively.The strain YAK11 was identified belonging to Neocallimastix frontalis by 18S rDNA sequence analyses. |
Key words: yak cellulolytic Hungate rolling tube technique 18S rDNA rumen anaerobic fungi |