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Biocatalyst for Pharmaceutical Industry

Author: YuanHongLing
Tutor: TaoWenYi
School: Jiangnan University
Course: Microbial and Biochemical Pharmacy
Keywords: Lipase Strain selection An organic solvent Fatty acid esters of ascorbic acid Biodiesel
CLC: R91
Type: Master's thesis
Year: 2006
Downloads: 268
Quote: 1
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Abstract


Lipase (EC3.1.1.3) is a lipid compounds are decomposed the synthesis and transesterification ring to the catalyst in the organic phase lipase can complete esterification, exchange and transesterification reaction regioselectivity, stereoselectivity, higher stability; inhibit the water in the side reactions, easy recovery and utilization; After completion of the reaction, separation and purification of the product is also relatively easy and can be completed using the chemical method difficult to carry out the racemic compound split, asymmetric synthesis, so that the an ancient enzyme species has a new and broad prospects for development. The enzyme reaction has incomparable advantages of the enzymatic reaction in the aqueous medium in a non-aqueous medium, it is more and more interested in the non-aqueous solution of the enzyme-catalyzed reaction. Lipase is widely distributed in the microbial world, but to seek high-yielding strains suitable for industrial production is very difficult, the domestic needs lipase still dependent on imports, in order to promote the non-aqueous phase enzyme-catalyzed research and application in our country, the need to develop lipases suitable for a non-aqueous phase varieties. The papers related to the activity of lipase in the organic solvent Producing Strain conditions for enzyme production and its application in the synthesis of ascorbic acid, fatty acid esters and biodiesel, the main results are as follows: (1) by adding 10 g / L of toluene pre-culture as the sole carbon source, and then from a soil sample in a transparent circle plate screening method successfully screened to an organic solvent-resistant lipase production by yeast A213, was identified as the yeast Yarrowia (of Yarrowia sp.) . (2) shake flask experiments show that the A213 suitable for enzyme production medium (g / L): yeast extract, 40, olive oil 10 the MgSO 4 · 7H 2 O KH 2 PO 4 optimal culture conditions for temperature 27 ° C, initial pH 6.5. Highest lipase activity up to 67.8 IU / mL; enzyme optimum temperature of 40 ° C, the optimum pH was 6.5, the enzyme in 40 ° C, 40 min when the enzyme activity can also keep 50%, the range of pH 5.5 to 8.5 Internal stability. Directly in tert-amyl alcohol solvent Catalytic Synthesis of L-ascorbyl palmitate. (3) for the first time reveals the fatty acid donor to grease the feasibility of synthesis of L-ascorbic acid fatty acid ester exchange reaction with the non-aqueous enzymatic ester of L-ascorbic acid; hydrogenated palm oil, soybean oil and seal oil fatty acid groups donor with the L-ascorbic acid ester interchange reaction medium were screened and found the highest concentration of product in t-amyl alcohol with the lipase-catalyzed reaction proceeds; discussed factors that affect the synthesis of ascorbic acid fatty acid ester, identified by optimum reaction conditions: optimum the initial fats substrate concentration is 200 to 600 mmol / L, and the optimal reaction time can be set for 9 h. Under optimum conditions, the product of concentration up to: 43.51 g / L. (4) fatty alcohol decomposition reaction is of great significance in the oils and fats industry. Established for the reaction product of qualitative and quantitative analysis of fatty alcohol solution by thin layer chromatography and high pressure liquid chromatography, Lipase-catalyzed without the addition of other solvents under the premise of hydrogenated palm oil fatty alcohol and anhydrous ethanol solution. process, when ethanol: palm oil = 200:1 reaction for 12 h, almost all converted into ethyl, palm oil, monoglycerides, the double Mannuronicate the content is close to 0.

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