Production of DagA, a ��-Agarase, by Streptomyces lividans in Glucose Medium or Mixed-Sugar Medium Simulating Microalgae Hydrolysate

Park, Ju Yi; Hong, Soon Kwang; Chang, Yong Keun

doi:10.4014/jmb.1405.05019

Cited by 10 publications

(7 citation statements)

References 23 publications

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“…S. lividans TK24/pUWL201-DagA was maintained on R2YE agar medium [17] containing thiostrepton (25 μg/mL). RSM3 broth (glucose, 2.5%; MgCl 2 ·6H 2 O, 0.5%; yeast extract, 1.1%; TES (C 6 H 15 NO 6 S, pH 7.2), 0.573%) which had been optimized for producing DagA from S. lividans TK24/pUWL201-DagA [6] was used for pre-cultivation (50 mL in 250-mL Erlenmeyer flask, 180 rpm, 60 h) and main fermentation (500 mL in 2-L Erlenmeyer flask, 200 rpm, 60 h) at 28 °C. The bacterial cell mass was removed by centrifugation at 10,000× g for 30 min at 4 °C, and the protein in the supernatant was concentrated by ammonium sulfate precipitation (85%).…”

Section: Methodsmentioning

confidence: 99%

“…The final filtrate was completely lyophilized, and the NAOs powder was stored at −20 °C under dark conditions and resuspended in sterile DW before use. The fine composition of the NAOs powder was analyzed by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) as described previously [6,16]. …”

Section: Methodsmentioning

confidence: 99%

“…HPLC (Waters Corporation, Milford, MA, USA) equipped with an evaporative light scattering detector (ELSD; Sedere, Alfortville, France) was used for the analysis [6]. Column, Asahipak NH2P-50 4E multi-mode column (250 × 4.6 mm); column temperature, 40 °C; mobile phase, acetonitrile:water (65:35); flow rate, 1 mL/min; ELSD detector nebulizer temperature, 50 °C.…”

Section: Figurementioning

confidence: 99%

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Anti-Obesity and Anti-Diabetic Effect of Neoagarooligosaccharides on High-Fat Diet-Induced Obesity in Mice

Hong

Lee²,

Kim³

et al. 2017

Marine Drugs

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Neoagarooligosaccharides (NAOs), mainly comprising neoagarotetraose and neoagarohexaose, were prepared by hydrolyzing agar with β-agarase DagA from Streptomyces coelicolor, and the anti-obesity and anti-diabetic effects of NAOs on high-fat diet (HFD)-induced obesity in mice were investigated after NAOs-supplementation for 64 days. Compared to the HFD group, the HFD-0.5 group that was fed with HFD + NAOs (0.5%, w/w) showed remarkable reduction of 36% for body weight gain and 37% for food efficiency ratios without abnormal clinical signs. Furthermore, fat accumulation in the liver and development of macrovesicular steatosis induced by HFD in the HFD-0.5 group were recovered nearly to the levels found in the normal diet (ND) group. NAOs intake could also effectively reduce the size (area) of adipocytes and tissue weight gain in the perirenal and epididymal adipose tissues. The increased concentrations of total cholesterol, triglyceride, and free fatty acid in serum of the HFD group were also markedly ameliorated to the levels found in serum of the ND group after NAOs-intake in a dose dependent manner. In addition, insulin resistance and glucose intolerance induced by HFD were distinctly improved, and adiponectin concentration in the blood was notably increased. All these results strongly suggest that intake of NAOs can effectively suppress obesity and obesity-related metabolic syndromes, such as hyperlipidemia, steatosis, insulin resistance, and glucose intolerance, by inducing production of adiponectin in the HFD-induced obese mice.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Figurementioning

confidence: 99%

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Anti-Obesity and Anti-Diabetic Effect of Neoagarooligosaccharides on High-Fat Diet-Induced Obesity in Mice

Hong

Lee²,

Kim³

et al. 2017

Marine Drugs

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“…Thus, in this study, we optimized the heterotrophic cultivation of promising strain of Chlorella sp. HS2 in order to explore the potential of green algae fermentation for biomass and biofuel production using Plackett-Burman design (PBD) 27 and the response surface methodology (RSM) with the central composite design (CCD) 26,28,29 . The optimized parameters were then experimentally verified on the flask scale and then on the final scale, a 5-L fermenter, for further optimization.…”

Section: Introductionmentioning

confidence: 99%

Optimization of heterotrophic cultivation of Chlorella sp. HS2 using screening, statistical assessment, and validation

Kim

Park

Lee

et al. 2019

Sci Rep

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The heterotrophic cultivation of microalgae has a number of notable advantages, which include allowing high culture density levels as well as enabling the production of biomass in consistent and predictable quantities. In this study, the full potential of Chlorella sp. HS2 is explored through optimization of the parameters for its heterotrophic cultivation. First, carbon and nitrogen sources were screened in PhotobioBox. Initial screening using the Plackett-Burman design (PBD) was then adopted and the concentrations of the major nutrients (glucose, sodium nitrate, and dipotassium phosphate) were optimized via response surface methodology (RSM) with a central composite design (CCD). Upon validation of the model via flask-scale cultivation, the optimized BG11 medium was found to result in a three-fold improvement in biomass amounts, from 5.85 to 18.13 g/L, in comparison to a non-optimized BG11 medium containing 72 g/L glucose. Scaling up the cultivation to a 5-L fermenter resulted in a greatly improved biomass concentration of 35.3 g/L owing to more efficient oxygenation of the culture. In addition, phosphorus feeding fermentation was employed in an effort to address early depletion of phosphate, and a maximum biomass concentration of 42.95 g/L was achieved, with biomass productivity of 5.37 g/L/D.

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“…The DagA activity assay was performed using the 3, 5‐dinitrosalicylic acid (DNS) method (Park et al ., 2014). After cultivation, the culture supernatant (2 ml) was collected by centrifugation at 13,000 rpm for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Enhanced production of neoagarobiose from agar with Corynebacterium glutamicum producing exo‐type and endo‐type β‐agarases

Jeon

Choi

Cho

et al. 2021

Microbial Biotechnology

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Neoagarobiose (NA2) derived from agar marine biomass is a rare reagent that acts as an antimelanogenesis reagent and moisturizer. Here, for the economical manufacturing of NA2, we developed the co-secretory production system of endo-type bagarases (DagA) and exo-type b-agarases (EXB3) in Corynebacterium glutamicum. For this purpose, we first developed a secretory system of DagA via Tat pathway. To improve the secretion efficiency, we coexpressed two Tat pathway components (TatA and TatC), and to improve the purity of secreted DagA in the culture supernatant, two endogenous protein genes (Cg2052 and Cg1514) were removed. Using the engineered strain (C. glutamicum SP002), we confirmed that DagA as high as 1.53 g l -1 was successfully produced in the culture media with high purity (72.7% in the supernatant protein fraction). Next, we constructed the expression system (pHCP-CgR-DagA-EXB3) for the simultaneous secretion of EXB3 via Sec-pathway together with DagA, and it was clearly confirmed that DagA and EXB3 were successfully secreted as high as 54% and 24.5%, respectively. Finally, using culture medium containing DagA and EXB3, we successfully demonstrated the conversion of high-concentration agar (40 g l -1 ) into NA2 via a two-stage hydrolysis process.

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Production of DagA, a ��-Agarase, by Streptomyces lividans in Glucose Medium or Mixed-Sugar Medium Simulating Microalgae Hydrolysate

Cited by 10 publications

References 23 publications

Anti-Obesity and Anti-Diabetic Effect of Neoagarooligosaccharides on High-Fat Diet-Induced Obesity in Mice

Anti-Obesity and Anti-Diabetic Effect of Neoagarooligosaccharides on High-Fat Diet-Induced Obesity in Mice

Optimization of heterotrophic cultivation of Chlorella sp. HS2 using screening, statistical assessment, and validation

Enhanced production of neoagarobiose from agar with Corynebacterium glutamicum producing exo‐type and endo‐type β‐agarases

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