Coenzyme Q10 (CoQ10), an important antioxidant molecule playing a major role in electron transport chain, has been commercially produced by fermentation process for the use in oral nutraceutical formulations. Construc...Coenzyme Q10 (CoQ10), an important antioxidant molecule playing a major role in electron transport chain, has been commercially produced by fermentation process for the use in oral nutraceutical formulations. Constructing the high-yielding CoQ10 producing strains is a pre-requisite for cost-effective production. A superior mutant strain P-87 generated from Paracoccus denitrificans ATCC 19367, which showed 1.25-fold improvement in specific CoQ10 content higher than the wild type strain at shake flask level, was selected to carry out the studies on CoQ10 yield improvement through fermenter process optimization. In the course of study, initially the cane-molasses-based medium and fed-batch fermentation strategy using pHBA in combination with sucrose were standardized in shake flask using wild type strain. This strategy was subsequently translated at 2 L laboratory fermenter while optimizing the fermentation process parameters using improved mutant strain P-87. Under optimized fermentation condition, mutant strain P-87 produced 49.85 mg/L of CoQ10 having specific content of 1.63 mg/g of DCW, which was 1.36 folds higher than the specific CoQ10 content of wild-type strain under similar optimized condition. The temperature and DO were found to be critical parameters for CoQ10 production by mutant strain P-87. The optimum temperature was found to be 32°C and the optimum DO concentration to be maintained throughout the fermentation cycle was found to be 30% of air saturation. Overall, a new cost-effective process has been established for the production of CoQ10 using the cheaper substrate “cane molasses” and higher CoQ10 producing mutant strain P-87.展开更多
2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Par...2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase(designated as Deh99), induced by DL-2-chloropropionate(DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography(Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant(Km) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40℃, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus.展开更多
Methomyl, an extremely toxic pesticide, is widely used in agriculture. A strain named mdw-1 capable of degrading methomyl rapidly was successfully isolated from activated sludge in this study. It could utilize methomy...Methomyl, an extremely toxic pesticide, is widely used in agriculture. A strain named mdw-1 capable of degrading methomyl rapidly was successfully isolated from activated sludge in this study. It could utilize methomyl as the sole carbon or nitrogen source. The optimal temperature and medium pH for its growth and methomyl biodegradation were 30℃ and 7.0, respectively. It was identified as a Paracoccus sp. according to its morphological features, physiological and biochemical characteristics, and phylogenetic analysis based on the sequence of 16S rDNA. Gas chromatography-mass spectrometry （GC-MS） analysis showed that methomyl could be completely transformed to S-methyl-N-hydroxythioacetamidate in 10 h of incubation with the isolate mdw-1.展开更多
As it was rear to find out detailed description, a study on biology and control measures ofP. marginatus was carried out at National Plant Quarantine Service, Katunayake Sri Lanka. Average length and width of differen...As it was rear to find out detailed description, a study on biology and control measures ofP. marginatus was carried out at National Plant Quarantine Service, Katunayake Sri Lanka. Average length and width of different stages, number of eggs in an egg sac, hatchability rate and duration of life cycle were studied. Control measures were tested using herbal oils and it was arranged in Completely Randomized Design with six replicates. Experiments were carried out in laboratory conditions under 28 ±2℃ and 70% RH. Range of length and width of different life stages revealed that, egg 0.3-0.1 mm ×0.15-0.10 mm, 1st instar 0.4-0.2 mm × 0.20-0.10 mm, 2nd instar 0.6-0.5 mm × 0.29-0.20 mm, 3rd instar male 0.8-0.5 mm × 0.30-0.20 mm, 3rd instar female 0.7-0.5 mm × 0.29-0.20 mm, adult male 0.9-0.7 mm× 0.20-0.10 mm and adult female 2.8-1.9 mm × 1.40-0.80 mm. A range of 100-200 eggs were in an ovisac and hatchability rate was 76-80%. Twenty to twenty-four days were taken to complete their life cycle. Cinnamon and Neem oil in cooperated with Surfactant and Kerosene oil could be effectively used as potential chemical agents for control of P. marginatus.展开更多
文摘Coenzyme Q10 (CoQ10), an important antioxidant molecule playing a major role in electron transport chain, has been commercially produced by fermentation process for the use in oral nutraceutical formulations. Constructing the high-yielding CoQ10 producing strains is a pre-requisite for cost-effective production. A superior mutant strain P-87 generated from Paracoccus denitrificans ATCC 19367, which showed 1.25-fold improvement in specific CoQ10 content higher than the wild type strain at shake flask level, was selected to carry out the studies on CoQ10 yield improvement through fermenter process optimization. In the course of study, initially the cane-molasses-based medium and fed-batch fermentation strategy using pHBA in combination with sucrose were standardized in shake flask using wild type strain. This strategy was subsequently translated at 2 L laboratory fermenter while optimizing the fermentation process parameters using improved mutant strain P-87. Under optimized fermentation condition, mutant strain P-87 produced 49.85 mg/L of CoQ10 having specific content of 1.63 mg/g of DCW, which was 1.36 folds higher than the specific CoQ10 content of wild-type strain under similar optimized condition. The temperature and DO were found to be critical parameters for CoQ10 production by mutant strain P-87. The optimum temperature was found to be 32°C and the optimum DO concentration to be maintained throughout the fermentation cycle was found to be 30% of air saturation. Overall, a new cost-effective process has been established for the production of CoQ10 using the cheaper substrate “cane molasses” and higher CoQ10 producing mutant strain P-87.
基金supported by National Basic Research Program of China(973 program,Grant No.2009CB724700)the Hundred Talent Program of the Chinese Academy of Sciences(A1097)National Natural Science Foundation of China(No.31100092)
文摘2-haloacid dehalogenases constitute a group of dehalogenases which are capable of dehalogenating the halogenated organic compounds. So far, the 2-haloacid dehalogenases have been found in many bacteria, but not in Paracoccus genus. In the present study, one enzyme 2-haloacid dehalogenase(designated as Deh99), induced by DL-2-chloropropionate(DL-2-CPA), was purified from the marine bacterium Paracoccus sp. DEH99, isolated from marine sponge Hymeniacidon perlevis. The enzyme of Deh99 was purified to homogeneity by ammonium sulfate precipitation, ion exchange chromatography(Q-Sepharose HP), and Superdex 200 gel filtration chromatography. The molecular weight of Deh99 was estimated to be 25.0 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE), and 50.0 kDa natively by gel filtration chromatography. The enzyme of Deh99 stereospecifically dehalogenated L-2-CPA to produce D-lactate, with an apparent Michaelis-Menten constant(Km) value of 0.21 mmol L-1 for L-2-CPA. The optimal pH and temperature for Deh99 activity were 10.0 and 40℃, respectively. The enzyme of Deh99 acted on short-carbon-chain 2-haloacids, with the highest activity towards monochloroacetate. The activity of Deh99 was slightly affected by DTT and EDTA, but strongly inhibited by Cu2+ and Zn2+. The enzyme of Deh99 shows unique substrate specificity and inhibitor sensitivities compared to previously characterized 2-haloacid dehalogenases and is the reported one about purified 2-haloacid dehalogenase isolated from the bacteria of Paracoccus genus.
基金Project supported by the National Natural Science Foundation of China(No.30600016)the National High Technology Research and Development Program of China(863 Program)(No.2007AA10Z405)+2 种基金the National Program for Sci-Tech Basic Platform Construction of China(No.2005DKA21201-2)the Program for Science and Technology Development ofJiangsu Province,China(No.BE2008669)the Program for Integration of Production,Education,and Research of Guangdong Province and Ministry of Education,China(No.2006090204007)
文摘Methomyl, an extremely toxic pesticide, is widely used in agriculture. A strain named mdw-1 capable of degrading methomyl rapidly was successfully isolated from activated sludge in this study. It could utilize methomyl as the sole carbon or nitrogen source. The optimal temperature and medium pH for its growth and methomyl biodegradation were 30℃ and 7.0, respectively. It was identified as a Paracoccus sp. according to its morphological features, physiological and biochemical characteristics, and phylogenetic analysis based on the sequence of 16S rDNA. Gas chromatography-mass spectrometry （GC-MS） analysis showed that methomyl could be completely transformed to S-methyl-N-hydroxythioacetamidate in 10 h of incubation with the isolate mdw-1.
文摘As it was rear to find out detailed description, a study on biology and control measures ofP. marginatus was carried out at National Plant Quarantine Service, Katunayake Sri Lanka. Average length and width of different stages, number of eggs in an egg sac, hatchability rate and duration of life cycle were studied. Control measures were tested using herbal oils and it was arranged in Completely Randomized Design with six replicates. Experiments were carried out in laboratory conditions under 28 ±2℃ and 70% RH. Range of length and width of different life stages revealed that, egg 0.3-0.1 mm ×0.15-0.10 mm, 1st instar 0.4-0.2 mm × 0.20-0.10 mm, 2nd instar 0.6-0.5 mm × 0.29-0.20 mm, 3rd instar male 0.8-0.5 mm × 0.30-0.20 mm, 3rd instar female 0.7-0.5 mm × 0.29-0.20 mm, adult male 0.9-0.7 mm× 0.20-0.10 mm and adult female 2.8-1.9 mm × 1.40-0.80 mm. A range of 100-200 eggs were in an ovisac and hatchability rate was 76-80%. Twenty to twenty-four days were taken to complete their life cycle. Cinnamon and Neem oil in cooperated with Surfactant and Kerosene oil could be effectively used as potential chemical agents for control of P. marginatus.
