Research On Trehalose Synthase Producing Methods

Trehalose owns wide prospect of application for its excellent features. After the invention of producing trehalose by enzyme catalysis, the trehalose production cost is greatly reduced and its commercial application becomes a reality. Compare to the double enzymatic conversion method, trehalose synthase can transform low-cost maltose into trehalose immediately, and thus become the research hotspot in the recent ten years.In search of heat resistant and high activity of trehalose synthetase, the thesis compared three different kinds of trehalose synthase production methods. First, the study screened the trehalose synthase producing bacteria, and found 1 strain, named L1204. The analysis shows that the produced trehalose synthetase has certain enzyme activities, but the activity is not high enough; The second way is starting from some available trehalose synthase producing strains, and try to build recombinant bacteria to produce more trehalose synthase. We bought two trains of bacteria from CGMCC and researched on trehalose synthase gene cloning and expression. While the Corynebacterium glutamicum showed the results as reported in literatures, the smegmatis bacteria did not show the expect results may due to materials, reagents, operation, or some other factors. Third, using modern bioinformatics means analysis those known gene sequences, and designed two gene sequences to synthesis and build gene engineering bacteria by the biotech companies. Then its enzymatic properties were verified, the ideal capable producing trehalose synthase engineered bacteria TSG2013 was obtained.Among above three trehalose synthase production methods, the work of screening trehalose synthase producing strains is huge workload and difficult, but the way is still an important way for getting new trehalose synthase. Using the available trehalose synthase producing strains to produce more trehalose synthase by gene cloning and expression, the relative workload greatly reduced and cost lower, but it need to be especially careful in actual operation of selecting bacteria materials, reagents and operating, otherwise it is difficult to get the desired results; About the last way, although the initial cost is higher, but it can greatly shorten the research time and get better results, therefore, it’s one of the most potential for the production of trehalose synthase.