Part I. 模擬移動床色譜法在人參皂苷、羥氯喹和蝦紅素分離中的應用 Part II. 靈芝和冬蟲夏草萃取物作為益生源提高益生菌維萊森斯芽孢桿菌抗菌活性試驗

Part I. Application of simulated moving bed chromatography to the separation of ginsenosides, hydroxychloroquine, and Astaxanthin Part II. A Test of Ganoderma lucidum and Cordyceps sinensis extracts as prebiotics to improve the antimicrobial activities of probiotic Bacillus velezensis

童珊珊(Tai Thi Ngoc Dong)

指導教授 ： 洪志勳

義守大學/工學院/化學工程學系暨生物技術與化學工程研究所/博士(2023年)

摘要

None

關鍵字

None

並列摘要

A two-part study was conducted with the applications of simulated moving bed (SMB) to separate ginsenosides, hydroxychloroquine and astaxanthin in the first part and evaluating the antibacterial activity of the probiotic Bacillus velezensis during stimulation of the prebiotics Ganoderma lucidium and Cordyceps sinensis in the second part. In the first part of this study, three different models of simulated moving bed chromatography were used, including reverse phase chromatography, chiral chromatography, and supercritical fluid chromatography. In batch chromatography, the physicochemical characteristics of separated materials and systems were determined by interacting with each other as stationary and mobile phases. Furthermore, triangle theory was applied to design parameters for the continuous mode. In the first reverse phase chromatography, the protopanaxadiol-type ginsenosides Rb1, Rc, and Rd from American ginseng were separated at 91.5 wt%; in the second separation, ginsenoside Rd was isolated at 89.5 wt%. The application involved chiral separation of the enantiomers of S-hydroxychloroquine from their racemic counterparts with remarkable purity and recovery values of 100% and 86.91%, respectively. Based on the results of the simulations from ASPEN, the experimental and simulated results are in reasonable agreement regarding the dispersion coefficients, mass transfer coefficients, and isotherm parameters. The optimal conditions were then identified for an increase in feed concentration of 10-fold and column length of 3.3-fold. By combining the configurations 2/3/3/2, solvent consumption was estimated to be reduced by 66%. Additionally, the use of supercritical fluid technology has also been applied to simulated moving bed chromatography (SF-SMB) in order to enhance elution power and cost-effectiveness. Based on the separation parameters set at ethanol 20 wt%, 40°C, and 140 bar, Astaxanthin was obtained with a purity of 96.44% and 90.02% recovery. As compared with liquid SMB, supercritical fluid produced 9.59 kg of feedstock per kg of stationary phase per day, approximately 72-fold higher in productivity.
The second part of study provides the feasibility of stimulating the growth and maintaining the viability of probiotic Bacillus velezensis strains BV1 and BV2. Probiotics cultured in BHI medium with Cordyceps sinensis (1-2 g L-1) or Ganoderma lucidum (2.25-4.5 g L-1) increased viability by 3.5-fold over the control. Among various cultured conditions, strain BV1 in addition to 2 g L-1 C. sinensis showed the highest antibacterial activity. The most well-defined inhibition zone was found against Salmonella spp. isolate S12 (20.13±0.56 mm), an approximate 18.94% increase in comparison to the control. Furthermore, E. coli demonstrated less impeded growth compared to Salmonella spp., representing strain D2-2 as the most susceptible among the strains investigated in this study, with an inhibition zone of 13.99±0.98 mm, 10.60% increase over the control. Probiotics were able to inhibit the growth of one out of four MRSA strains with growth turbidity despite ineffectiveness against Klebsiella pneumoniae and Acinetobacter baumannii. As part of the screening process, the optimal pH was measured to be 4 to 8, with survival rates of 85.89 to 96.66% for strain BV1 and 82.12 to 96.00% for strain BV2. For improved temperature tolerance, sporulation-forming was carried out on potato dextrose agar medium supplemented with 1% sodium chloride, calcium chloride, and potassium chloride. Based on the high sporulation efficiency of approximately 93% in the BV1 strain and the consideration of an economical cost for further scale-up, the use of 1% w/v NaCl resulted a 1.7-fold increase in temperature tolerance (80oC for 30 min) compared to the vegetative strain. As a result, this study presents a feasible methodology for mass production of probiotics and an alternative to antibiotics by including prebiotics like C. sinensis (2 g L-1) and a method for spore-forming with a stable shelf life for a growing market.

並列關鍵字

SMB ； SF-SMB ； Astaxanthin ； ginsenosides ； hydroxychloroquine ； separation ； probiotic ； antimicrobial activity ； Sporulation ； Bacillus velezensis

參考文獻

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