Reported here is a protocol to fabricate a biocatalyst with high enzyme loading and activity retention, from the conjugation of electrospun nanofibrous membrane having biomimetic phospholipid moiety and lipase. To imp...Reported here is a protocol to fabricate a biocatalyst with high enzyme loading and activity retention, from the conjugation of electrospun nanofibrous membrane having biomimetic phospholipid moiety and lipase. To improve the catalytic efficiency and activity of the immobilized enzyme, poly（acrylonitrile-co-2-methacryloyloxyethyl phosphorylcholine）s（PANCMPCs） were, respectively, electrospun into nanofibrous membranes with a mean diameter of 90 nm, as a support for enzyme immobilization. Lipase from Candida rugosa was immobilized on these nanofibrous membranes by adsorption. Properties of immobilized lipase on PANCMPC nanofibrous membranes were compared with those of the lipase immobilized on the polyacrylonitrile（PAN） nanofibrous and sheet membranes, respectively. Effective enzyme loading on the nanofibrous membranes was achieved up to 22.0 mg/g, which was over 10 times that on the sheet membrane. The activity retention of immobilized lipase increased from 56.4% to 76.8% with an increase in phospholipid moiety from 0 to 9.6%（molar fraction） in the nanofibrous membrane. Kinetic parameter Km was also determined for free and immobilized lipase. The Km value of the immobilized lipase on the nanofibrous membrane was obviously lower than that on the sheet membrane. The optimum pH was 7.7 for free lipase, but shifted to 8.3-8.5 for immobilized lipases. The optimum temperature was determined to be 35 ℃ for the free enzyme, but 42-44℃ for the immobilized ones, respectively. In addition, the thermal stability, reusability, and storage stability of the immobilized lipase were obviously improved compared to the free one.展开更多
Lipase from Candida rugosa was covalently immobilized on the surface of an uhrafihration hollow fiber membrane fabricated from poly （ acrylonitrile-co-maleic acid） （ PANCMA ） in which the carboxyl groups were acti...Lipase from Candida rugosa was covalently immobilized on the surface of an uhrafihration hollow fiber membrane fabricated from poly （ acrylonitrile-co-maleic acid） （ PANCMA ） in which the carboxyl groups were activated with 1-ethyl-3-（ dimethylaminopropyl ） carbodiimide hydrochloride （ EDC ） and dicyclohexyl carbodiimide （ DCC ）/ N-hydroxyl succinimide（NHS）, respectively. The properties of the immobilized lipase were assayed and compared with those of the free enzyme. The maximum activities were observed in a relatively broader pH value range at high temperatures for the immobilized lipase compared to the free one. It was also found that the thermal and pH stabilities of lipase were improved upon immobilization and at 50 ℃ the thermal inactivation rate constant values are 2. 1 × 10^ -2 for the free lipase, 3.2 × 10^-3 for the immobilized lipase on the EDC-activated PANCMA membrane and 3.5 × 10^-3 for the immobilized lipase on the DCC/NHS-activated PANCMA membrane, respectively.展开更多
PVA/ Chitosan (CS) composite membrane was studied in this paper, which could be used for enzyme processing of fat and oils. The parameters such as concentration of lipase, pH, and cross-linking agent as well as metal ...PVA/ Chitosan (CS) composite membrane was studied in this paper, which could be used for enzyme processing of fat and oils. The parameters such as concentration of lipase, pH, and cross-linking agent as well as metal ions, which influence the immobilization of lipase in membrane, were optimized. The immobilized lipase was 0.66 u/cm2 and the recovery of immobilized lipase activity was 24%. The membrane reactor could be used to synthesize monoglyceride (MG) with many batches.展开更多
PVA/Chitosan(CS) composite membrane was used for enzyme processing of fats and oils. The concentration of lipase and cross-linking agent which influence the immobilization of lipase in membrane were determined. Epichl...PVA/Chitosan(CS) composite membrane was used for enzyme processing of fats and oils. The concentration of lipase and cross-linking agent which influence the immobilization of lipase in membrane were determined. Epichlorohydrin is used as the cross-linking agent. The immobilized lipase is 0.66 u·cm -2 and the recovery of immobilized lipase is 24%. The membrane reactor was tested to synthesis monoglyceride(MG), which could be used many times without loss conversion yield of MG. The PVA/CS lipase membrane reactor is a new reactor for lipase catalytic biphase systems.展开更多
A multiphase enzyme membrane reactor using aqueous organic biphase instead of water phase alone as the reaction medium was employed to investigate the lipase catalyzed synthesis of bioactive dipeptides. The medium eff...A multiphase enzyme membrane reactor using aqueous organic biphase instead of water phase alone as the reaction medium was employed to investigate the lipase catalyzed synthesis of bioactive dipeptides. The medium effect on dipeptide yield was first studied. When N acetyl L phenylalanine ethyl ester(APEE) was used as a carboxyl component, the reactivity order of amino acid amides was found to be L Leu NH 2> L Val NH 2> L Ala NH 2> L Gly NH 2. The didpetide, N Ac L Phe L Leu NH 2, could be synthesized in the multiphase enzyme membrane reactor in a high yield and purity due to the simultaneous separation and reaction.展开更多
基金Supported by the National Natural Science Foundation of China for Distinguished Young Scholars(No50625309)the National Postdoctoral Science Foundation of China(No20060400337)
文摘Reported here is a protocol to fabricate a biocatalyst with high enzyme loading and activity retention, from the conjugation of electrospun nanofibrous membrane having biomimetic phospholipid moiety and lipase. To improve the catalytic efficiency and activity of the immobilized enzyme, poly（acrylonitrile-co-2-methacryloyloxyethyl phosphorylcholine）s（PANCMPCs） were, respectively, electrospun into nanofibrous membranes with a mean diameter of 90 nm, as a support for enzyme immobilization. Lipase from Candida rugosa was immobilized on these nanofibrous membranes by adsorption. Properties of immobilized lipase on PANCMPC nanofibrous membranes were compared with those of the lipase immobilized on the polyacrylonitrile（PAN） nanofibrous and sheet membranes, respectively. Effective enzyme loading on the nanofibrous membranes was achieved up to 22.0 mg/g, which was over 10 times that on the sheet membrane. The activity retention of immobilized lipase increased from 56.4% to 76.8% with an increase in phospholipid moiety from 0 to 9.6%（molar fraction） in the nanofibrous membrane. Kinetic parameter Km was also determined for free and immobilized lipase. The Km value of the immobilized lipase on the nanofibrous membrane was obviously lower than that on the sheet membrane. The optimum pH was 7.7 for free lipase, but shifted to 8.3-8.5 for immobilized lipases. The optimum temperature was determined to be 35 ℃ for the free enzyme, but 42-44℃ for the immobilized ones, respectively. In addition, the thermal stability, reusability, and storage stability of the immobilized lipase were obviously improved compared to the free one.
文摘Lipase from Candida rugosa was covalently immobilized on the surface of an uhrafihration hollow fiber membrane fabricated from poly （ acrylonitrile-co-maleic acid） （ PANCMA ） in which the carboxyl groups were activated with 1-ethyl-3-（ dimethylaminopropyl ） carbodiimide hydrochloride （ EDC ） and dicyclohexyl carbodiimide （ DCC ）/ N-hydroxyl succinimide（NHS）, respectively. The properties of the immobilized lipase were assayed and compared with those of the free enzyme. The maximum activities were observed in a relatively broader pH value range at high temperatures for the immobilized lipase compared to the free one. It was also found that the thermal and pH stabilities of lipase were improved upon immobilization and at 50 ℃ the thermal inactivation rate constant values are 2. 1 × 10^ -2 for the free lipase, 3.2 × 10^-3 for the immobilized lipase on the EDC-activated PANCMA membrane and 3.5 × 10^-3 for the immobilized lipase on the DCC/NHS-activated PANCMA membrane, respectively.
文摘PVA/ Chitosan (CS) composite membrane was studied in this paper, which could be used for enzyme processing of fat and oils. The parameters such as concentration of lipase, pH, and cross-linking agent as well as metal ions, which influence the immobilization of lipase in membrane, were optimized. The immobilized lipase was 0.66 u/cm2 and the recovery of immobilized lipase activity was 24%. The membrane reactor could be used to synthesize monoglyceride (MG) with many batches.
文摘用吸附法固定脂肪酶时,膜材料的亲疏水性对固定化酶的量、比活力和活力稳定性等有很大影响.今以柱状假丝酵母脂肪酶和猪胰脂肪酶为研究对象,选取了8种亲疏水性不同的膜材料（醋酸纤维素、聚丙烯腈、聚酰胺、聚砜、聚醚砜、聚偏二氟乙烯、聚丙烯和聚四氟乙烯）作为固定化载体,用吸附法制备了固定化脂肪酶膜.研究结果表明,强疏水性聚四氟乙烯和聚丙烯膜对两种酶的吸附量都比较大,且固定化酶的比活力和活力回收率比较高,聚四氟乙烯固定化柱状假丝酵母酶比游离态酶的半衰期提高了6倍以上.强亲水性醋酸纤维素膜对猪胰脂肪酶的吸附量比聚四氟乙烯高,但是固定化酶的比活力、活力回收率比强疏水性膜低,而接触角在40°～50°的聚酰胺膜和聚砜膜的吸附量最小.因此吸附法制备固定化脂肪酶膜,选择聚丙烯膜和聚四氟乙烯膜是合适的,制备的优化条件为吸附温度25℃,酶溶液的pH为7.5,吸附时间10 h.
基金国家自然科学基金! (No .2 96760 0 1 )山东大学国家微生物技术重点实验室开放基金河北省自然科学基金资助
文摘PVA/Chitosan(CS) composite membrane was used for enzyme processing of fats and oils. The concentration of lipase and cross-linking agent which influence the immobilization of lipase in membrane were determined. Epichlorohydrin is used as the cross-linking agent. The immobilized lipase is 0.66 u·cm -2 and the recovery of immobilized lipase is 24%. The membrane reactor was tested to synthesis monoglyceride(MG), which could be used many times without loss conversion yield of MG. The PVA/CS lipase membrane reactor is a new reactor for lipase catalytic biphase systems.
文摘A multiphase enzyme membrane reactor using aqueous organic biphase instead of water phase alone as the reaction medium was employed to investigate the lipase catalyzed synthesis of bioactive dipeptides. The medium effect on dipeptide yield was first studied. When N acetyl L phenylalanine ethyl ester(APEE) was used as a carboxyl component, the reactivity order of amino acid amides was found to be L Leu NH 2> L Val NH 2> L Ala NH 2> L Gly NH 2. The didpetide, N Ac L Phe L Leu NH 2, could be synthesized in the multiphase enzyme membrane reactor in a high yield and purity due to the simultaneous separation and reaction.
