Bacillus mycoides TKU039 發酵烏賊軟骨所生產幾丁聚醣酶之分離與定性

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Title:	Bacillus mycoides TKU039 發酵烏賊軟骨所生產幾丁聚醣酶之分離與定性
Other Titles:	Isolation and characteristic of chitosanases produced by Bacillus mycoides TKU039 fermented with squid pen
Authors:	陳俊谷;Chen, Chun-Ku
Contributors:	淡江大學化學學系碩士班王三郎;Wang, San-Lang
Keywords:	幾丁聚醣酶;烏賊軟骨粉;幾丁寡糖;Bacillus mycoides;chitosanase;squid pen powder;oligomer
Date:	2016
Issue Date:	2017-08-24 23:38:38 (UTC+8)
Abstract:	實驗菌株TKU039 係篩選自新北市林口山區紅土之幾丁聚醣酶生產菌，屬於格蘭氏陽性，蕈狀孢芽桿菌，經由16S rDNA 部分鹼基序列分析與NCBI/BLAST資料庫比對，得知此菌株最接近Bacillus 屬，經由API鑑定系統判定為Bacillus mycoides，鑑定率為88.5 %。以含有1%烏賊軟骨粉、0.1% K2HPO4、0.05% MgSO4 ·7H2O之150 mL液態培養基，於37°C中搖瓶(150 rpm)培養5天可得較高幾丁聚醣酶活性(0.65U)。依較佳培養條件培養所得之發酵液，經離心、硫酸銨沉澱、DEAE-Sepharose及Macro-Prep® DEAE Cartridge陰離子交換層析等步驟，純化度(61.7倍)以及活性回收率(4.2%)，純化後經SDS PAGE電泳測得分子量約為41 kDa之幾丁聚醣酶。此幾丁聚醣酶的最適反應pH為6、pH安定性為pH4~pH10 、最適反應溫度為50°C、於50°C以下皆具熱安定性。此幾丁聚醣酶水解WSC後之寡醣經過MALDI-TOF-MS分析得知其組成多為3~6醣，並以6醣含量最多。 Chitosan has been more popular in recent years due to its good bio-compatibility, low toxicity and rarely no allergic reaction to human body. Chitosan also has other bioactivity like reduce blood pressure, lower cholesterol, increase immunity, and its antibacterial property can be used in many clinical applications. For the best resource of Chitin/Chitosan we extracted from the seafood wastes (squid pens, shrimp or crab shells), hich is abundant of polysaccharides, and glucosamine that we can use enzyme hydrolysis to achieve it. In traditional way, using a lot of chemical to receive chitosan and its oligomer, but in nowadays, we use the natural way, chitosanase bacteria strain TKU039 identified as Bacillus sp. from the soil in New Taipei City Linkou mountain. TKU039 was culture in medium with the squid pen powder (SPP) and shrimp powder (SP).The SPP medium was found most suitable with 1.0% SPP as a sole carbon/nitrogen source optimized under cultured at 37°C with 150 rpm for 5 days in 150 mL of medium containing 1% SPP, 0.1% K2HPO4 and 0.05 % MgSO4·7H2O.
Appears in Collections:	[Graduate Institute & Department of Chemistry] Thesis

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