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Dihydro lipo a mid e dehydrog en as e catalyzes the oxidation of dihydrolipoamide: dihydrolipoamide + NAD+ -> lipoamide + NADH + H+. In case of bacteria and eucarya, this enzyme normally functions as an integral component of the pyruvate, 2-oxoglutarate, and branched chain 2-oxoacid dehydrogenase multienzyme complex or the glycine cleavage system. A novel 1368-bp Clostridium kluyveri gene codes for ~ 50 kDa protein was cloned and over expressed in E.coli system and further biochemical and spectral studies were sought to investigate its identity. The recombinant protein showed very strong dihydrolipoamide dehydrogenase and diaphorase activity. Non-covalantly attached FAD was released at boiling temperature and quantified (7.2M). UV-visible absorption spectra showed two peaks at 453nm and 270nm and the ratio of A280/450 = 4.98. Although the optimum temperature was 40ºC at pH 7.0, the protein showed some stability as high as 70ºC and 6.0-9.0 pH range. Fluorescent spectral study revealed the formation of NADH in forward reaction in presence of reduced lipoamide, in turn confirmed dihydrolipoamide dehydrogenase activity. Unlike other anaerobic bacterial dihydrolipoamide dehydrogenase enzyme, this enzyme had specificity to only NADH as coenzyme, where NADPH did not react at all.

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Loss of SHA1 function showed uninucleate and 4C single guard cells where plastid number are similar with WT paired guard cells stoma confirming that the mutation blocked guard mother