Proteus vulgaris에서 유래한 리파아제의 고정화 및 바이오디젤 생산
Production of Biodiesel Using Immobilized Lipase from Proteus vulgaris.
1
조회
0
다운로드
2
인용
0
참고문헌
키워드
초록
바이오디젤은 긴 사슬 지방산의 알킬 에스테르로서 동물성지방 또는 식물성 오일과 알코올이 반응하여 에스테르 교환반응에 의해 생성되는 대체연료이다. 지난 십여 년 동안, 다양한 리파아제를 이용한 바이오디젤 생산에 대해 연구되었다. 하지만 효소 촉매 공정을 통한 바이오디젤 생산의 경우,높은 효소 단가로 산업적 공정에 쉽게 적용할 수 없었다. 이러한 문제점을 극복하기 위해, 저렴한 오일 원료를 선택하거나, 바이오디젤 생산에 적합한 리파아제를 스크리닝하는 과정또는 리파아제 고정화 방법이 활발히 연구되었다. 이번 연구에서는 P. vulgaris에서 유래한 리파아제 K80을 E. coli균에서발현하여 얻은 효소액으로 바이오디젤을 생산하였다. 재조합리파아제 K80은 높은 발현량을 보였으며, 높은 가수분해 반응의 비활성도(specific activity)와 유기용매에서 높은 안정성을 확인했다. 리파아제 K80은 올리브 오일과 메탄올을 3-stepwise 방법을 이용하여 바이오디젤을 생산할 수 있었다. 리파아제 K80을 소수성 결합을 이용하여 담체 표면에 흡착시켜 얻은 고정화 K80을 이용하여 수용성 리파아제 K80과 동일한 방법으로 바이오디젤을 생산한 결과, 효율적으로 바이오디젤 생산을 확인했다. 고정화 K80은 다양한 식물성 오일과메탄올을 사용하여 효과적으로 바이오디젤을 생산하였다. 고정화 K80을 이용하여 바이오디젤 생산뿐만 아니라 다른 산업적 공정에서도 활용할 수 있을 것으로 기대한다.
Abstract
Biodiesel, mono-alkyl esters of long chain fatty acids, is one of the alternative fuels derived from renewable lipid feedstock, such as vegetable oils or animal fats. For decade, various lipases have been used for the production of biodiesel. However, the production of biodiesel by enzymatic catalyst has profound restriction in industry application due to high cost. To overcome these problems, many research groups have studied extensively on the selection of cheap oil sources, the screening of suitable lipases, and development of lipase immobilization methods. In this study, we produced biodiesel from plant oil using Proteus vulgaris lipase K80expressed in Escherichia coli cells. The recombinant lipase K80 was not only expressed in high level but also had high specific lipase activity and high stability in various organic solvents. Lipase K80 could produce biodiesel from olive oil by 3-stepwise methanol feeding method. The immobilized lipase K80 also produced biodiesel using the same 3-stepwise method. The immobilized lipase could produce biodiesel efficiently from various plant oils and waste oils.
참고문헌 (24)
- Ackman, R. G., "The “basic” fatty acid composition of Atlantic fish oils:Potential similarities useful for enrichment of polyunsaturatedfatty acids by urea complexation", J. Am. Oil Chem. Soc, 65, pp.136~138 (1988)
- Akoh, C. C., "Enzymatic approach to biodiesel production", J. Agric. FoodChem, 55, pp.8995~9005 (2007)
- Bajaj, A., "Biodiesel production through lipase catalyzed transesterification:an overview", J. Mol. Catal. B Enzym, 62, pp.9~14 (2010)
- Charpe, T. W, "Biodiesel productionusing waste frying oil", Waste Manag, 31, pp.85~90 (2010)
- Cunha, A. G., "Immobilization of Yarrowia lipolyticalipase - a comparison of stability of physical adsorptionand covalent attachment techniques", Appl. Biochem. Biotechnol, 146, pp.49~56 (2007)
- Dizge, N., "Biodieselproduction from canola oil by using lipase immobilizedonto hydrophobic microporous styrene-divinylbenzenecopolymer", Biochem. Eng. J, 44, pp.220~225 (2009)
- Du, W., "Comparativestudy on lipase-catalyzed transformation of soybean oilfor biodiesel production with different acyl acceptors", J.Mol. Catal. B Enzym, 30, pp.125~129 (2004)
- Jaeger, K. E, "Lipases for biotechnology", Curr. Opin. Biotechnol, 13, pp.390~397 (2002)
- Jegannathan, K. R., "Production of biodiesel using immobilizedlipase - a critical review", Crit. Rev. Biotechnol, 28, pp.253~264 (2008)
- Kharrat, N., "Immobilization of Rhizopus oryzaelipase on silica aerogels by adsorption: Comparison with thefree enzyme", Process Biochem, 46, pp.1083~1089 (2011)
- Kim, H. K., "Characterization of an alkaline lipase from Proteus vulgarisK80 and the DNA sequence of the encoding gene", FEMSMicrobiol. Lett, 135, pp.117~121 (1996)
- Kim, H. K., "Partialinterfacial activation of Proteus vulgaris lipase overexpressedin Escherichia coli", Biosci. Biotechnol. Biochem, 60, pp.1365~1367 (1996)
- Lee, H. W., "Overexpression of an alkaline lipasegene from Proteus vulgaris K80 in Escherichia coli BL21", pKLE, 2000, pp.22, 1543~1547
- Li, N. W., "Highly efficienttransformation of waste oil to biodiesel by immobilizedlipase from Penicillium expansum", Process Biochem, 44, pp.658~688 (2009)
- Lu, J., "Pretreatment of immobilized Candida sp.99-125 lipaseto improve its methanol tolerance for biodiesel production", J.Mol. Catal. B Enzym, 62, pp.15~18 (2010)
- Parawira,W, "Biotechnological production of biodieselfuel using biocatalyzed transesterification: A review", Crit.Rev. Biotechnol, 29, pp.82~93 (2009)
- Salis, A., "Comparison among immobilized lipases on macroporouspolypropylene toward biodiesel synthesis", J. Mol. Catal. BEnzym, 54, pp.19~26 (2008)
- Shimada, Y., "Conversion ofvegetable oil to biodiesel using immobilized Candidaantarctica lipase", J. Am. Oil Chem. Soc, 76, pp.789~793 (1999)
- Shimada, Y., "Enzymatic alcoholysis for biodiesel fuel productionand application of the reaction to oil processing", J. Mol.Catal. B Enzym, 17, pp.133~142 (2002)
- Tan, T., "Biodieselproduction with immobilized lipase: A review", Biotechnol.adv, 28, pp.628~634 (2010)
- Tongboriboon, K., "Mixed lipases for efficient enzymatic synthesis of biodieselfrom used palm oil and ethanol in a solvent-free system", J.Mol. Catal. B Enzym, 67, pp.52~59 (2010)
- Wang, Y., "Immobilized recombinant Rhizopus oryzaelipase for the production of biodiesel in solvent free system", J. Mol. Catal. B Enzym, 67, pp.45~51 (2010)
- Yang, K. S., "Catalytic properties of a lipase from Photo bacterium lipolyticum for biodiesel production containing a high methanol concentration", J. Biosci. Bioeng, 107(6), pp.599~604 (2009)
- Yoo, H. Y., "novel alkalinelipase from Ralstonia with potential application in biodieselproduction", Bioresour. Technol, 102, pp.6104~6111 (2011)