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A thermostable recombinant transaldolase with high activity over a broad pH range
Song-Yan Huang1, Y-H Percival Zhang, Jian-Jiang Zhong
1State Key Laboratory of Bioreactor Engineering, School of Bioengineering, East China University of Science and Technology, Shanghai, China.
Applied Microbiology and Biotechnology
|September 28, 2011
Summary
Researchers discovered a highly thermostable transaldolase (TAL) from Thermotoga maritima, active at 80°C. This enzyme shows potential for biofuel production and carbohydrate synthesis applications.
Area of Science:
- Biochemistry
- Enzymology
- Biotechnology
Background:
- Thermophilic enzymes are crucial for industrial applications due to their stability at high temperatures.
- Transaldolase (TAL) is an important enzyme in carbohydrate metabolism.
- Identifying novel thermostable enzymes is key for advancing biotechnological processes.
Purpose of the Study:
- To clone and express the transaldolase (TAL) gene (TM0295) from the hyper-thermophilic bacterium Thermotoga maritima.
- To characterize the enzymatic activity and thermostability of the recombinant T. maritima transaldolase at elevated temperatures.
- To explore the potential applications of this ultra-thermostable enzyme.
Main Methods:
- Cloning and expression of the TM0295 gene in Escherichia coli.
- Enzyme activity assays at various temperatures and pH levels.
- Thermostability determination through half-life measurements at 60 °C and 80 °C.
- Calculation of the total turnover number at 80 °C.
Main Results:
- The recombinant transaldolase from T. maritima was successfully expressed.
- The enzyme demonstrated significant activity at 80 °C, a first for transaldolase.
- Exceptional thermostability was observed, with half-lives of 198 hours at 60 °C and 13.0 hours at 80 °C.
- A high turnover number of 1.5 × 10^6 mol product/mol enzyme at 80 °C was estimated.
- The enzyme maintained high activity across a broad pH range (6.0-9.0).
Conclusions:
- The T. maritima transaldolase is an ultra-thermostable enzyme with high catalytic activity at elevated temperatures.
- This enzyme exhibits potential for industrial applications, including enzymatic biofuel production and cell-free synthesis of high-value carbohydrates.
- The findings present a promising biocatalyst for high-temperature biotransformations.
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