台灣本土溫泉嗜熱菌的調查、收集、鑑定及其農業上的應用(III)─台灣本土溫泉嗜熱菌的調查、收集、鑑定及其農業上的應用(III)
Date Issued
2004-07-31
Date
2004-07-31
Author(s)
張文章
DOI
922317B002027
Abstract
1. Isolation of thermophilic bacteria and construction of strain library: We have surveyed 65
hot springs in Taiwan to collect and analyze the bacteria contained therein. The bacterial strains
were isolated, purified, cultured and identified. Up to date we have obtained over 1000 bacterial
strains. These are deposited in Academia Sinica, National Taiwan University and Tzu-Chi
University. These bacteria were identified by Gram stain, electron microscopy and 16S rDNA
sequencing and enzymatic analysis. They were found to belong to Bacillus sp.， Geobacillus sp.，
Alicyclobacillus sp.， Brevibacillus sp.， Deinococcus sp.， Rubrobacter sp., Paracraurococcu
sp., Thermus sp.， Meiothermus sp.， Rhodothermus sp.， Chloroflexus sp.， Roseiflexus sp.,
Thermonema sp. and Pseudoxanthomonas sp.
2. Enzymatic assay: Analyses of the enzymatic properties revealed that 20 % of the thermophilic
strains contain amylase activity, 25 % contain protease activity and 38 % contain lipase activity.
These strains are being analyzed in order to find bacteria with higher enzymatic activity.
3. Thermostable aminotransferases: Four aminotransferases were selected for study, including
aminotransferase (AspAT), phenylalanine aminotransferase (PheAT), branched-chain amino acid
aminotransferase (BCAT) and D-amino acids aminotransferase (DAT). In preliminary studies,
Deinococcus taiwanensis 407T and D. taiwanensis 539 with highly enzymatic activities of AspAT
and BCAT. Pseudoxanthomonas taiwanensis CB-225has highly enzymatic activities of AspAT
and PheAT.
4. We have isolated several new strains with remarkable resistance to UV and
gamma-radiation, such as Rubrobacter taiwanensis LS-293T and Deinococcus taiwanensis
WR-407T. By using high dose of UV and gamma-radiation to treat the thermophilic bacteria we
also found 87 strains of thermophilic and radiation-resistant bacteria, which can grow at
temperatures from 50~70 °C. By 16S rDNA sequencing we found some of these bacteria to
belong to Deinococcus sp., Rubrobacter sp., Bacillus sp., Geobacillus sp., Thermus sp.
5. Gene cloning of thermophilic enzymes and construction of a shuttle vector: We have
screened 53 bacterial strains which contain plasmids and constructed a shuttle vector by using the
plasmid existing in NTU-103. This shuttle vector will be useful for the construction of expression
vector for the large-scale expression of thermophilic enzymes.
Publisher
臺北市：國立臺灣大學生化科學研究所
Type
report
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