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Thermoclostridium stercorarium subsp. stercorarium DSM 8532

ABSTRACT: Clostridium stercorarium stercorarium BW, DSM 8532 Genome sequencing

PROVIDER: PRJNA181307 | ENA |

REPOSITORIES: ENA

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Thermoclostridium stercorarium subsp. stercorarium DSM 8532

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Heterologous Expression and Characterization of a Thermostable α-L-Rhamnosidase from <i>Thermoclostridium stercorarium</i> subsp. <i>thermolacticum</i> DSM 2910 and Its Application in the Biotransformation of Rutin.

Project description:An α-L-rhamnosidase gene from Thermoclostridium. stercorarium subsp. thermolacticum DSM 2910 (TstRhaA) was cloned and expressed. The maximum TstRhaA activity of the protein reached 25.2 U/ml, and the molecular mass was approximately 106.6 kDa. The protein was purified 8.0-fold by Ni-TED affinity with an overall recovery of 16.6% and a specific activity of 187.9 U/mg. TstRhaA activity was the highest at 65°C and pH 6.5. In addition, it exhibited excellent thermal stability, better pH stability, good tolerance to low concentrations of organic reagents, and high catalytic activity for p-nitrophenyl-α-L-rhamnopyranoside (pNPR). Substrate specificity studies showed that TstRhaA exhibited a high specific activity for rutin. At 60°C, pH 6.5, and 0.3 U/ml enzyme dosage, 60 g/l rutin was converted to 45.55 g/l isoquercitrin within 150 min. The molar conversion rate of rutin and the yield of isoquercitrin were 99.8% and 12.22 g/l/h, respectively. The results suggested that TstRhaA could be used for mass production of isoquercitrin.

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