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메틸프룩토시드를 이용한 과당계열 당 지방산 에스테르의 효소적 합성
허주형,김해성 한국생물공학회 1998 KSBB Journal Vol.13 No.6
과당계열 배당체인 메틸프룩토시드를 반응출발문질로하여 과당계열 당 지방산 에스테르인 메틸프룩토시드 올레산 모노 및 디에스테르를 효소적 용매 및 무용매공정에 의하여 합성하였다. 용매공정으로부터 머틸프룩토시드 올레산 모노에스테르를 합성한 결과, 최적 상용용매로는 2-메틸-2-프로관올이 우수하게 나타났으며 매틸프룩토시드의 용해도는 $60^{\circ}C$애서 400g/L이상에 상당하였다. 최적반응조건을 검토한 결과, 에틸프룩토시드 올레산의 최적올비 3.1, 에틸프룩토시드의 최적 초기농도 200g/L, Novozym 435의 최적 촉매함량 1%(w!v), 반응온도 $60^{\circ}C$에서 반응시간 8시간만에 70%의 평 형 젠화율애 도달하였으며 12.6g/L-hr의 생산성으로 매렐프룩토시드 올레산 모노에스테프 블 생싼할 수 았었다. 그리고, 무용매공정에서 메틸프룩토시드 올레산 디에스터르를 합생한 결과, 메틸프룩토시드와 올레산의 최적돌에 l.2, Novozym 435의 최적 촉매함령, 10%(w/w), 반응온도 $70^{\circ}C$, 진공도 20 - 200mmHg 에서 반응시간 10시간만에 95%이상의 전화율로 140g;L-hr의 생산성으로 메틸프룩토시드 올레산 디에스테르를 생산할 수 있었다. Enzymatic synthesis of fructose-based sugar fatty acid esters, such as methyl fructoside oleic acid mono and diester, was investigated using methyl fructoside as a sugar starting material. For the production of methyl fructoside fatty acid monoester by solvent process, 2-methyl 2-propanol was found to be a god reaction medium resulting a higher yield and productivity due to its high sugar solubility. The yield and productivity of methyl fructoside oleic acid monoester were 70% and 12.6g/L-hr, respectively, when molar ratio of methyl fructoside, initial concentration of methyl fructoside, enzyme(Novozym 435) content, and reaction temperature were 3:1, 200g/L, 1%(w/v), and $60^{\circ}C$, respectively. Methyl fructoside oleic acid diester was prepared by lipase-catalyzed diacylation of methyl fructoside and oleic acid in the solvent-free process. Maximum yield of 98% and productivity of 140g/L-hr were achieved when molar ratio(methyl fructoside and oleic acid) of 1:2 enzyme content of 10%(w/v) and reaction temperature of $70^{\circ}C$ were applied for the operating conditions under a reduced pressure of 20∼200 mmHg.
Methyl- $\beta$ -D-Fructofuranoside 합성을 위한 고정화 전화당 효소의 미소환경 최적화
허주형,안형환 대한안전경영과학회 1999 대한안전경영과학회지 Vol.1 No.1
In order to enhance the selectivity, productivity and yield of methyl fructoside, which was synthesized by enzymatic glycosylation of sucrose and methanol solution, controlling of surface property of solid support using different immobilization procedures optimized microenvironment of immobilized invertase. Silanization and polyethylene imine coating methods were adopted to give a hydrophobic and hydrophilic environment of immobilized invertase. As a result, polyethyleneimine coating method gave higher loading of enzyme, effective activity, and relative activity than silanization method, because it brought on increasing the functional density of amino group and enhancing the conservation of activity by regulating of hydrophilicity. And then, hydrophilic environment was possible to restraint the assessing of methyl fructoside molecule, which was more hydrophobic than sucrose, fructose, and glucose molecule in the reaction mixture, into .the active site of immobilizedinvertase. Consequently, hydrophilic microenvironment of immobilized invertase by polyethyleneimine coating obtained higher yield and productivity with increasing conversion than silanized and native invertase. Thus, this procedure optimized the microenvironment of immobilized invertase suitable for the enzymatic synthesis of methyl fructoside.