KR20110002951A - Method for preparing hydroxyalkanoate alkylester using the microorganisms producing polyhydroxyakanoate - Google Patents

Method for preparing hydroxyalkanoate alkylester using the microorganisms producing polyhydroxyakanoate Download PDF

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KR20110002951A
KR20110002951A KR1020090060496A KR20090060496A KR20110002951A KR 20110002951 A KR20110002951 A KR 20110002951A KR 1020090060496 A KR1020090060496 A KR 1020090060496A KR 20090060496 A KR20090060496 A KR 20090060496A KR 20110002951 A KR20110002951 A KR 20110002951A
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hydroxybutyrate
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이상엽
박진환
정유경
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한국과학기술원
바이오퓨얼켐 주식회사
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Abstract

PURPOSE: A method for preparing HA alkyl ester using microorganisms with PHA(polyhydroxyalkyanoate) production is provided to be used in the industrial production of 3-hydroxybutyrate alkyl ester(3-HB alkyl ester). CONSTITUTION: A method for preparing hydroxyalkanoate alkyl ester comprises: a step of culturing microorganisms with polyhydroxyalkanoate production to obtain polyhydroxyalkanoate; a step of performing autolysis of polyhydroxyalkanoate in the microorganisms to obtain hydroxyalkanoate; and a step of adding alcohol and reacting to obtain hydroxyalkanoate alkylester.

Description

폴리하이드록시알카노에이트 생성능을 가지는 미생물을 이용한 하이드록시알카노에이트 알킬에스테르의 제조방법{Method for Preparing hydroxyalkanoate alkylester Using the Microorganisms Producing polyhydroxyakanoate}Method for preparing hydroxyalkanoate alkylester using the microorganisms Producing polyhydroxyakanoate

본 발명은 폴리하이드록시알카노에이트(polyhydroxyalkanoate;PHA) 생성능을 가지는 미생물을 이용한 하이드록시알카노에이트 알킬에스테르(HA alkylester)의 제조방법에 관한 것으로서, 더욱 상세하게는 PHA 생성능을 가지는 미생물을 배양하여, PHA를 대량생산하고, PHA를 포함하는 배양액의 pH를 조절하여 하이드록시알카노에이트(HA) 단량체를 제조한 뒤, 알킬에스테르화시키는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르(HA alkylester)의 제조방법에 관한 것이다. The present invention relates to a method for preparing hydroxyalkanoate alkyl ester (HA alkylester) using a microorganism having a polyhydroxyalkanoate (PHA) generating ability, and more particularly, by culturing a microorganism having a PHA generating ability , Hydroxyalkanoate alkyl ester (HA alkylester), characterized in that the mass production of PHA, by adjusting the pH of the culture medium containing PHA to produce a hydroxyalkanoate (HA) monomer, alkyl esterification It relates to a manufacturing method.

최근 고유가와 환경 문제로 인해 미생물을 이용한 바이오연료 생산이 큰 관심을 끌고 있다. 최근 바이오 디젤이 경유를 대체하거나 경유와 혼합하여 디젤엔진에 사용할 수 있는 대체연료로 부상하면서 시장 규모가 매우 빠른 속도로 증가하고 있다. 유럽연합(EU)에서는 2008년 한 해 동안 660만톤의 바이오디젤이 생산 되었으며, 5.5 billion 유로의 시장규모를 이루었다 (Biodiesel Market, Frost & Sullivan). 또한, 미국에서도 2006년 한 해 동안 3억 갤론의 바이오디젤이 생산되었다 (Biodiesel Market, Global Industry Analysts Inc, 2006. 5). 바이오디젤은 연소율이 좋아 유독가스의 배출이 낮고, 경유보다 발열량이 10% 정도 낮으며, 발화점은 더 높아 운송과 저장에 보다 안전하다는 장점을 지니고 있다. 바이오디젤은 주로 동식물의 지방성분을 경유와 유사한 특성을 갖도록 가공하거나, 식물성 유지 (쌀겨, 폐식용유, 대두유, 유채유 등)와 알코올을 반응시켜 제조하였다. 이 경우 대량생산이 어렵다는 단점이 있다. 따라서 경유의 대체연료로 적합한 바이오디젤을 미생물을 이용하여 대량 생산한다면, 원유 수입 대체 효과 및 온실 가스 배출 감소로 인한 환경적 효과 등을 가져올 수 있다. Recently, due to high oil prices and environmental problems, biofuel production using microorganisms has attracted great attention. Recently, the market size is increasing at a very rapid pace, with biodiesel emerging as a substitute fuel that can be used for diesel engines as a substitute for or mixed with diesel. In 2008, the EU produced 6.6 million tonnes of biodiesel, with a market size of 5.5 billion euros (Biodiesel Market, Frost & Sullivan). In the United States, 300 million gallons of biodiesel were produced in 2006 alone (Biodiesel Market, Global Industry Analysts Inc, May 2006). Biodiesel has the advantages of high combustion rate, low emission of toxic gas, 10% lower calorific value than diesel, and higher flash point, making it safer for transportation and storage. Biodiesel is mainly prepared by processing the fat component of animals and plants to have properties similar to those of diesel, or by reacting vegetable oils (rice bran, waste cooking oil, soybean oil, rapeseed oil, etc.) with alcohol. In this case, mass production is difficult. Therefore, if biodiesel suitable for diesel fuel is mass-produced using microorganisms, it can bring about the effects of importing crude oil and environmental effects from reducing greenhouse gas emissions.

PHA는 미생물이 탄소원은 풍부하나 다른 성장인자(질소, 인, 산소, 황 등)가 부조할 때 세포내에서 합성하여 축적하는 에너지 저장물질이며, 성장환경이 바뀌어 제한되었던 성장인자가 다시 제공되면 축적해 놓은 PHA를 분해하여 에너지원으로 사용하게 된다. PHA는 생산하는 미생물의 종류나, 공급되는 화학물질의 종류, 배양조건 등에 변화에 의해 100종류 이상의 단량체가 구성요서로 가능한 것으로 알려져 있다 (Steinbchel and Valentin, FEMS Microbiol Lett, 128:219, 1995).PHA is an energy storage substance synthesized and accumulated in cells when microorganisms are rich in carbon sources but other growth factors (nitrogen, phosphorus, oxygen, sulfur, etc.) help. The PHA is broken down and used as an energy source. PHA is known to be composed of more than 100 types of monomers by the type of microorganisms produced, the type of chemicals supplied, the culture conditions (Steinbchel and Valentin, FEMS Microbiol Lett, 128: 219, 1995).

최근, 미생물이 생산하는 PHA를 메틸에스테르화하여, 바이오디젤을 생산하려는 노력들이 진행되어오고 있으며, 정제된 PHA에 황산과 메탄올을 가하여 고온고압조건에서 반응시켜 3-하이드록시알카노에이트 메틸에스테르(3-HA methylester)를 생산한 기술이 개발된바 있다(Zhang, Xiaoujun et al.,Biomacromolecules, 10:707, 2009).Recently, efforts have been made to produce biodiesel by methyl esterifying PHA produced by microorganisms, adding sulfuric acid and methanol to purified PHA and reacting under high temperature and high pressure to obtain 3-hydroxyalkanoate methyl ester ( A technique for producing 3-HA methylester has been developed (Zhang, Xiaoujun et al . , Biomacromolecules , 10: 707, 2009).

그러나, 상기 기술은 PHB(poly-R-3-hydroxybutyrate)에서 R-3하이드록시부티레이트 메틸에스테르로의 전환율이 52%에 불과하며, 생산되는 R-3하이드록시부티레이트 메틸에스테르의 순도가 저하되는 단점이 있다.However, the above technique has a drawback of only 52% conversion of poly-R-3-hydroxybutyrate (PHB) to R-3 hydroxybutyrate methyl ester, and lowered purity of the produced R-3 hydroxybutyrate methyl ester. There is this.

이에, 본 발명자들은 미생물을 이용하여 바이오디젤을 생산하는 새로운 방법을 개발하고자 예의 노력한 결과, Alcaligenes latus 균주를 이용하여 poly(3-hydroxybutyrate)를 생산하고, 이를 균주 내에서 자가분해시킨후, 알킬에스테르화하는 경우, 고효율로 3-HB alkylester를 생산할 수 있다는 것을 확인하고, 본 발명을 완성하게 되었다.Therefore, the present inventors have made diligent efforts to develop a new method for producing biodiesel using microorganisms. As a result, poly (3-hydroxybutyrate) is produced using Alcaligenes latus strain, and it is self-decomposed in the strain, followed by alkyl ester. In this case, it was confirmed that 3-HB alkylester can be produced with high efficiency, and the present invention was completed.

결국, 본 발명의 목적은 폴리하이드록시알카노에이트 생산균주를 이용하여, in vivo에서 자가분해시켜, 하이드록시알카노에이트 단량체를 제조하는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르를 고효율로 제조하는 방법을 제공하는데 있다.As a result, an object of the present invention is to produce a hydroxyalkanoate alkyl ester having high efficiency by self-decomposing in vivo using a polyhydroxyalkanoate producing strain to produce a hydroxyalkanoate monomer. To provide a way.

상기 목적을 달성하기 위하여, 본 발명은 (a) 폴리하이드록시알카노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알카노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알카노에이트를 함유하는 미생물에서 폴리하이드록시알카노에이트를 자가분해시켜, 하이드록시알카노에이트을 제조하는 단계; 및 (c) 상기 제조된 하이드록시알카노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알카노에이 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알카노에이트 알킬에스테르의 제조방법을 제공한다. In order to achieve the above object, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxyalkanoate producing ability, to produce a polyhydroxyalkanoate; (b) self-degrading polyhydroxyalkanoate in the microorganism containing polyhydroxyalkanoate to produce hydroxyalkanoate; And (c) adding alcohol to the hydroxyalkanoate prepared above, and reacting to prepare a hydroxyalkanoate alkylester.

또한, 본 발명은 (a) 폴리하이드록시알케노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알케노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알케노에이트를 함유하는 미생물에서 폴리하이드록시알케노에이트를 자가분해시켜, 하이드록시알케노에이트를 제조하는 단계; 및 (c) 상기 제조된 하이드록시알케노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알케노에이트 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알케노에이트 알킬에스테르의 제조방법을 제공한다.In addition, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxy alkenoate production ability, to produce a polyhydroxy alkenoate; (b) self-degrading polyhydroxyalkenoate in the microorganism containing polyhydroxyalkenoate to prepare hydroxyalkenoate; And (c) adding an alcohol to the prepared hydroxyalkenoate and reacting to prepare a hydroxyalkenoate alkylester.

또한, 본 발명은 (a) 폴리하이드록시알키노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알키노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알키노에이트를 함유하는 미생물에서 폴리하이드록시알키노에이트를 자가분해시켜, 하이드록시알키노에이트를 제조하는 단계; 및 (c) 상기 제조된 하이드록시알키노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알키노에이트 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알키노에이트 알킬에스테르의 제조방법을 제공한다.In addition, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxyalkynoate production ability, to produce a polyhydroxyalkynoate; (b) self-degrading polyhydroxyalkynoate in the microorganism containing polyhydroxyalkynoate to prepare hydroxyalkynoate; And (c) adding an alcohol to the prepared hydroxyalkynoate and reacting to prepare a hydroxyalkynoate alkylester.

본 발명은 또한, (a) 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성능을 가지는 미생물을 배양하여, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}를 생성시키는 단계; (b) 상기 생성된 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}을 함유하는 미생물에서 폴리-3-하이드록시부티레이트을 자가분해시켜, 3-하이드록시부티레이트를 제조하는 단계; 및 (c) 상기 3-하이드록시부티레이트에 메탄올을 첨가하고, 반응시켜 3-하이드록시부티레이트 메틸에스테르를 제조하는 단계를 포함하는 3-하이드록시부티레이트 메틸에스테르의 제조방법을 제공한다.The present invention also comprises the steps of (a) culturing a microorganism having the ability to produce poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)}, to produce poly-3-hydroxybutyrate (poly (3-hydroxybutyrate)} ; (b) self-degrading poly-3-hydroxybutyrate in a microorganism containing the produced poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)} to prepare 3-hydroxybutyrate; And (c) adding methanol to the 3-hydroxybutyrate and reacting to prepare 3-hydroxybutyrate methyl ester.

본 발명에 따르면, 미생물이 생산하는 poly(3-hydroxybutyrate)를 100% 전환 효율로 3-하이드록시부티레이트 알킬에스테르(3-HB alkylester)로 전환할 수 있어, 최근 바이오디젤로의 효용가치가 확인된 3-하이드록시부티레이트 알킬에스테르의 산업적 생산 방법으로 유용하다.According to the present invention, it is possible to convert poly (3-hydroxybutyrate) produced by microorganisms to 3-hydroxybutyrate alkylester (3-HB alkylester) at 100% conversion efficiency, and thus the utility value of biodiesel has recently been confirmed. It is useful as an industrial production method of 3-hydroxybutyrate alkyl ester.

일 관점에서, 본 발명은 (a) 폴리하이드록시알카노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알카노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알카노에이트를 함유하는 미생물에서 폴리하이드록시알카노에이트를 자가분해시켜, 하이드록시알카노에이트을 제조하는 단계; 및 (c) 상기 제조된 하이드록시알카노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알카노에이트 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알카노에이트 알킬에스테르의 제조방법에 관한 것이다.In one aspect, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxyalkanoate generating ability, to produce a polyhydroxyalkanoate; (b) self-degrading polyhydroxyalkanoate in the microorganism containing polyhydroxyalkanoate to produce hydroxyalkanoate; And (c) adding an alcohol to the prepared hydroxyalkanoate and reacting to prepare a hydroxyalkanoate alkylester.

본 발명에 있어서, 하이드록시알카노에이트는 방향족 고리기, 에폭시기, 시아노기, 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것을 사용할 수 있으나, 이에 한정되는 것은 아니다. In the present invention, hydroxyalkanoate may be modified with a substituent selected from the group consisting of an aromatic ring group, an epoxy group, a cyano group, a halogen group, but is not limited thereto.

본 발명에서, 폴리하이드록시알카노에이트의 자가분해는 세포내에 축적된 폴리하이드록시알카노에이트가 세포내 분해효소에 의하여 자가분해되는 것을 의미하며, 통상 폴리하이드록시알카노에이트의 자가분해는 균체를 pH 2~12의 조건에서 정치하였을 경우에 수행하며, 바람직하게는, 배양액의 pH를 2~5로 조절하고, 34~50℃에서 정치반응시켜 수행할 수 있다.In the present invention, self-decomposition of polyhydroxyalkanoate means that the polyhydroxyalkanoate accumulated in the cell is autolyzed by the intracellular degrading enzyme, and the autolysis of the polyhydroxyalkanoate is usually a cell Is performed when the mixture is left at a pH of 2 to 12, and preferably, the pH of the culture solution is adjusted to 2 to 5, and the reaction may be performed at 34 to 50 ° C.

본 발명의 일 양태에서, Alcaligenes latus의 경우, pH 3~4에서 반응온도 37℃에서 30분~10시간 정치반응하였을 때, 가장 높은 자가분해능을 나타내었다.In one embodiment of the present invention, Alcaligenes latus showed the highest self-decomposition when left standing for 30 minutes to 10 hours at a reaction temperature of 37 ° C. at pH 3-4.

본 발명에 있어서, 상기 (c) 단계는 유기용매를 추가로 첨가하여 수행하는 것을 특징으로 할 수 있으며, 바람직하게는 클로로포름을 첨가하여 수행할 수 있다.In the present invention, the step (c) may be performed by additionally adding an organic solvent, and preferably, may be performed by adding chloroform.

본 발명에 있어서, 폴리하이드록시알카노에이트 생성능을 가지는 미생물은 예를 들면, Aeromonas 속 미생물, Achromobacter 속 미생물, Acidovorax delafieldii, Acidovax facilis,, Acinetobacter 속 미생물, Actinomyces 속 미생물, Aeromonas 속 미생물, Alcaligenes 속 미생물, Alteromonas 속 미생물, Amoebobacter 속 미생물, Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp., Azospirillum 속 미생물, Azotobacter 속 미생물, Bacillus 속 미생물, Beggiatoa 속 미생물, Beijerinckia 속 미생물, Beneckea 속 미생물, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter 속 미생물, Chlorogloea 속 미생물, Chromatium 속 미생물, Chromobacterium 속 미생물, Clostridium 속 미생물, Comamonas 속 미생물, Corynebacterium 속 미생물, Cyanobacteria 속 미생물, Derxia 속 미생물, Desulfonema 속 미생물, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira 속 미생물, Ferrobacillus ferroxidans, Flavobacterium sp., Haemophilus influenzae, Halobacterium 속 미생물, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, Hydrogenophaga 속 미생물, Hyphomicrobium 속 미생물, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., Leptothrix discophorus, Methylobacterium 속 미생물, Methylosinus 속 미생물, Micrococcus 속 미생물, Mycobacterium 속 미생물, Nitrobacter 속 미생물, Nocardia 속 미생물, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium 속 미생물, Physarum ploycephalum, Pseudomonas 속 미생물, Ralstonia 속 미생물, Rhizobium 속 미생물, Rhodobacillus 속 미생물, Rhodobacter 속 미생물, Rhodococcus 속 미생물, Rhodocyclus 속 미생물, Rhodomicrobium vannielii, Rhodopseudomonas 속 미생물, Rhodospirillum 속 미생물, Sphingomonas paucimobilis, Spirillum 속 미생물, Spirulina 속 미생물, Staphylococcus 속 미생물, Stella 속 미생물, Streptomyces 속 미생물, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus 속 미생물, Thiocapsa 속 미생물, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea 속, 폴리하이드록시알카노에이트 생성능을 가지는 효소를 코딩하는 유전자로 형질전환된 미생물을 들 수 있다.In the present invention, the microorganism having the ability to produce polyhydroxyalkanoate is, for example, the genus Aeromonas , Achromobacter microorganisms, Acidovorax delafieldii , Acidovax facilis , Microorganisms of the genus Acinetobacter , Actinomyces microorganisms, Microorganisms of the genus Aeromonas , Microorganisms of the genus Alcaligenes , Alteromonas genus microorganism, Amoebobacter genus microorganism, Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp ., Azospirillum spp, Azotobacter spp, Bacillus spp, Beggiatoa spp, Beijerinckia spp, Beneckea spp, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter spp, Chlorogloea spp ., Chromatium microorganism of the genus, Chromobacterium spp, Clostridium spp, Comamonas spp, Corynebacterium spp, Cyanobacteria in microorganisms, Derxia spp, Desulfonema spp, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira spp, Ferrobacillus ferroxidans, Flavobacterium sp, Haemophilus influenzae, Halobacterium spp, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, microorganisms in Hydrogenophaga, microorganisms in Hyphomicrobium, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., L eptothrix discophorus, Methylobacterium spp, Methylosinus spp, Micrococcus spp, Mycobacterium spp, Nitrobacter spp, Nocardia spp, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium spp, Physarum ploycephalum, Pseudomonas spp, Ralstonia spp , Rhizobium spp, Rhodobacillus spp, Rhodobacter spp, Rhodococcus spp, Rhodocyclus spp, Rhodomicrobium vannielii, Rhodopseudomonas spp, Rhodospirillum spp, Sphingomonas paucimobilis, Spirillum spp, Spirulina spp, Staphylococcus spp, Stella spp , Streptomyces spp, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus spp, Thiocapsa spp, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea in And microorganisms transformed with a gene encoding an enzyme having a polyhydroxyalkanoate generating ability.

본 발명에 있어서, 상기 하이드록시알카노에이트는 락테이트(lactate), 2-하이드록시부탄산(hydroxybutyrate), 3-하이드록시프로피온산(hydroxypropionate), 3-하이드록시부티레이트(hydroxybutyrate), 3-하이드록시발레르산(hydroxyvalerate), 4-하이드록시부탄산(hydroxybutyrate), 탄소수가 6~14개인 중간사슬 길이의 (D)-3-하이드록시카르복실산(hydroxycarboxylic acids), 3-하이드록시프로피온산(hydroxypropionic acid), 3-하이드록시헥산산(hydroxyhexanoic acid), 3-하이드록시헵탄산(hydroxyheptanoic acid), 3-하이드록시옥탄산(hydroxyoctanoic acid), 3-하이드록시노난산(hydroxynonanoic acid), 3-하이드록시데칸산(hydroxydecanoic acid), 3-하이드록시운데칸산(hydroxyundecanoic acid), 3-하이드록시 도데칸산(hydroxydodecanoic acid), 3-하이드록시테트라데칸산(hydroxytetradecanoic acid), 3-하이드록시헥사데칸산(hydroxyhexadecanoic acid), 4-하이드록시발레르산(hydroxyvaleric acid), 4-하이드록시헥산산(hydroxyhexanoic acid), 4-하이드록시헵탄산(hydroxyheptanoic acid), 4-하이드록시옥탄산(hydroxyoctanoic acid), 4-하이드록시데칸산(hydroxydecanoic acid), 5-하이드록시발레르산(hydroxyvaleric acid), 5-하이드록시헥산산(hydroxyhexanoic acid), 6-하이드록시도데칸산(hydroxydodecanoic acid), 3-하이드록시(hydroxy)-4-펜텐산(pentenoic acid), 3-하이드록시(hydroxy)-4-trans-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-4-cis-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-5-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-6-trans-옥텐산(octenoic acid), 3-하이드록시(hydroxy)-6-cis-옥텐산(octenoic acid), 3-하이드록시(hydroxy)-7-옥텐산(octenoic acid), 3-하이드록시(hydroxy)-8-노넨산(nonenoic acid), 3-하이드록시(hydroxy)-9-데센산(decenoic acid), 3-하이드록시(hydroxy)-5-cis-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-6-cis-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-5-cis-테트라데센산(tetradecenoic acid), 3-하이드록 시(hydroxy)-7-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-5,8-cis-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-4-메틸발레르산(methylvaleric acid), 3-하이드록시(hydroxy)-4-메틸헥산산(methylhexanoic acid), 3-하이드록시(hydroxy)-5-메틸헥산산(methylhexanoic acid), 3-하이드록시(hydroxy)-6-메틸헵탄산(methylheptanoic acid), 3-하이드록시(hydroxy)-4-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-5-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-6-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-7-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-6-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-7-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-8-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-7-메틸데칸산(methyldecanoic acid), 3-하이드록시(hydroxy)-9-메틸데칸산(methyldecanoic acid), 3-하이드록시(hydroxy)-7-메틸-6-옥텐산(octenoic acid), 말산(malic acid), 3-하이드록시숙신산(hydroxysuccinic acid)-메틸에스테르, 3-하이드록시아디핀산(hydroxyadipinic acid)-메틸에스테르, 3-하이드록시스베린산(hydroxysuberic acid)-메틸에스테르, 3-하이드록시아젤라인산(hydroxyazelaic acid)-메틸에스테르, 3-하이드록시세바신산(hydroxysebacic acid,)-메틸에스테르, 3-하이드록시스베린산(hydroxysuberic acid)-에틸에스테르, 3-하이드록시세바신산(hydroxysebacic acid)-에틸에스테르, 3-하이드록시피메린산(hydroxypimelic acid)-프로필에스테르, 3-하이드록시세바신산(hydroxysebacic acid)-벤질에스테르, 3-하이드록시(hydroxy)-8-아세톡시옥탄산(acetoxyoctanoic acid), 3-하이드록시(hydroxy)-9-아세톡시노난산(acetoxynonanoic acid), 페녹시(phenoxy)-3-하이드록시부티레이트(hydroxybutyric acid), 페녹시(phenoxy)-3-하이드록시발레르산(hydroxyvaleric acid), 페녹시(phenoxy)-3-하이드록시헵탄산(hydroxyheptanoic acid), 페녹시(phenoxy)-3-하이드록시옥탄산(hydroxyoctanoic acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시부티레이트(hydroxybutyric acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시발레르산(hydroxyvaleric acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시헥산산(hydroxyhexanoic acid), para-니트로페녹시(nitrophenoxy)-3-하이드록시헥산산(hydroxyhexanoic acid), 3-하이드록시(hydroxy)-5-페닐발레르산(phenylvaleric acid), 3-하이드록시(hydroxy)-6-페닐헥산(phenylhexanoic acid), 3-하이드록시(hydroxy)-7-페닐헵탄산(phenylheptanoic acid), 3-하이드록시(hydroxy)-8-페닐옥탄산(phenyloctanoic acid), 3-하이드록시(hydroxy)-9-페닐노난산(phenylnonanoic acid), 3-하이드록시(hydroxy)-10-페닐데칸산(phenyldecanoic acid), 3-하이드록시(hydroxy)-5-시클로헥실부탄산(cyclohexylbutyric acid), 3-하이드록시(hydroxy)-5-시클로헥실부탄산(cyclohexylbutyric acid), 3,12-디하이드록시도데칸산(dihydroxydodecanoic acid), 3,8-디하이드록시(dihydroxy)-5-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-4,5-에폭시데칸산(epoxydecanoic acid), 3-하이드록시(hydroxy)-6,7-에폭시도데칸산(epoxydodecanoic acid), 3-하이드록시(hydroxy)-8,9-에폭시(epoxy)-5,6-cis-테트라데칸산(tetradecanoic acid), 7-시아노(cyano)-3-하이드록시헵탄산(hydroxyheptanoic acid), 9-시아노(cyano)-3-하이드록시노난 산(hydroxynonanoic acid), 3-하이드록시(hydroxy)-7-플루오로헵탄산(fluoroheptanoic acid), 3-하이드록시(hydroxy)-9-플루오로노난산(fluorononanoic acid), 3-하이드록시(hydroxy)-6-클로로헥산산(chlorohexanoic acid), 3-하이드록시(hydroxy)-8-클로로옥탄산(chlorooctanoic acid), 3-하이드록시(hydroxy)-6-브로모헥산산(bromohexanoic acid), 3-하이드록시(hydroxy)-8-브로모옥탄산(bromooctanoic acid), 3-하이드록시(hydroxy)-11-브로모운데칸산(bromoundecanoic acid), 3-하이드록시(hydroxy)-2-부텐산(butenoic acid), 6-하이드록시(hydroxy)-3-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-2-메틸부탄산(methylbutyric acid), 3-하이드록시(hydroxy)-2-메틸발레르산(methylvaleric acid) 및 3-하이드록시(hydroxy)-2,6-디메틸-5-헵텐산(heptenoic acid)로 구성된 군 (Steinbㆌchel, A and Valentin, H. E., FEMS Microbiology Letters, 128: 219-228, 1995)에서 선택된 하나 이상인 것을 특징으로 할 수 있으나, 이에 한정되는 것은 아니다.In the present invention, the hydroxyalkanoate is lactate, 2-hydroxybutyrate, 3-hydroxypropionate, 3-hydroxybutyrate, 3-hydroxy Valeric acid, 4-hydroxybutyrate, medium chain length (D) -3-hydroxycarboxylic acids having 6 to 14 carbon atoms, 3-hydroxypropionic acid ), 3-hydroxyhexanoic acid, 3-hydroxyheptanoic acid, 3-hydroxyoctanoic acid, 3-hydroxynonanoic acid, 3-hydroxy Decanoic acid, 3-hydroxyundecanoic acid, 3-hydroxydodecanoic acid, 3-hydroxytetradecanoic acid, 3-hydroxyhexadecanoic acid acid), 4-hydr Hydroxyvaleric acid, 4-hydroxyhexanoic acid, 4-hydroxyheptanoic acid, 4-hydroxyoctanoic acid, 4-hydroxydecanoic acid ), 5-hydroxyvaleric acid, 5-hydroxyhexanoic acid, 6-hydroxydodecanoic acid, 3-hydroxy-4-pentenoic acid (pentenoic) acid, 3-hydroxy-4-trans-hexenoic acid, 3-hydroxy-4-cis-hexenoic acid, 3-hydroxy-5 -Hexenoic acid, 3-hydroxy-6-trans-octenic acid, 3-hydroxy-6-cis-octenic acid, 3-hydroxy (hydroxy) -7-octenoic acid, 3-hydroxy-8-nonenoic acid, 3-hydroxy-9-decenoic acid, 3-hydroxy Hydroxy-5-cis-dodecenoic acid, 3-hydroxy xy) -6-cis-dodecenoic acid, 3-hydroxy-5-cis-tetradecenoic acid, 3-hydroxy-7-cis-tetradecenoic acid (tetradecenoic acid), 3-hydroxy-5,8-cis-cis-tetradecenoic acid, 3-hydroxy-4-methylvaleric acid, 3-hydroxy Hydroxy-4-methylhexanoic acid, 3-hydroxy-5-methylhexanoic acid, 3-hydroxy-6-methylheptanoic acid , 3-hydroxy-4-methyloctanoic acid, 3-hydroxy-5-methyloctanoic acid, 3-hydroxy-6-methyloctanoic acid (methyloctanoic acid), 3-hydroxy-7-methyloctanoic acid, 3-hydroxy-6-methylnonanoic acid, 3-hydroxy-7 Methylnonanoic acid, 3-hydroxy-8-methylnonanoic acid acid, 3-hydroxy-7-methyldecanoic acid, 3-hydroxy-9-methyldecanoic acid, 3-hydroxy-7-methyl acid 6-octennoic acid, malic acid, 3-hydroxysuccinic acid-methyl ester, 3-hydroxyadipinic acid-methyl ester, 3-hydroxysveric acid (hydroxysuberic acid) -methyl ester, 3-hydroxyazelaic acid-methyl ester, 3-hydroxysebacic acid, -methyl ester, 3-hydroxysuberic acid-ethyl Ester, 3-hydroxysebacic acid-ethyl ester, 3-hydroxypimelic acid-propyl ester, 3-hydroxysebacic acid-benzyl ester, 3-hydroxy ( hydroxy-8-acetoxyoctanoic acid, 3-hydroxy-9-acetoxynonanoic acid xynonanoic acid, phenoxy-3-hydroxybutyric acid, phenoxy-3-hydroxyvaleric acid, phenoxy-3-hydroxyheptanoic acid ( hydroxyheptanoic acid, phenoxy-3-hydroxyoctanoic acid, para-cyanophenoxy-3-hydroxybutyric acid, para-cyanophenoxy -3-hydroxyvaleric acid, para-cyanophenoxy-3-hydroxyhexanoic acid, para-nitrophenoxy-3-hydroxyhexanoic acid), 3-hydroxy-5-phenylvaleric acid, 3-hydroxy-6-phenylhexane, 3-hydroxy-7-phenylheptane Phenylheptanoic acid, 3-hydroxy-8-phenyloctanoic acid, 3-hydroxy-9-phenylnonanoic acid, 3- Hydroxy-10-phenyldecanoic acid, 3-hydroxy-5-cyclohexylbutyric acid, 3-hydroxy-5-cyclohexylbutanoic acid ( cyclohexylbutyric acid), 3,12-dihydroxydodecanoic acid, 3,8-dihydroxy-5-cis-tetradecenoic acid, 3-hydroxy- 4,5-epoxydecanoic acid, 3-hydroxy-6,7-epoxydodecanoic acid, 3-hydroxy-8,9-epoxy- 5,6-cis-tetradecanoic acid, 7-cyano-3-hydroxyheptanoic acid, 9-cyano-3-hydroxynonanoic acid ), 3-hydroxy-7-fluoroheptanoic acid, 3-hydroxy-9-fluorononanoic acid, 3-hydroxy-6- Chlorohexanoic acid, 3-hydroxy-8-chloro Chlorooctanoic acid, 3-hydroxy-6-bromohexanoic acid, 3-hydroxy-8-bromooctanoic acid, 3-hydroxy )-11-bromooundecanoic acid, 3-hydroxy-2-butenoic acid, 6-hydroxy-3-dodecenoic acid, 3-hydroxy Hydroxy-2-methylbutyric acid, 3-hydroxy-2-methylvaleric acid and 3-hydroxy-2,6-dimethyl-5-hep One or more selected from the group consisting of heptenoic acid (Steinb ㆌ chel, A and Valentin, HE, FEMS Microbiology Letters , 128: 219-228, 1995), but is not limited thereto.

본 발명에 있어서, 상기 (a) 단계의 배양은 질소원이 제한된 배지에서 수행하는 것을 특징으로 할 수 있으며, 상기 (d) 단계의 반응은 80~120℃에서 1~24시간 수행하는 것을 특징으로 할 수 있다.In the present invention, the step (a) may be characterized in that the nitrogen source is carried out in a restricted medium, the reaction of step (d) is characterized in that it is carried out for 1 to 24 hours at 80 ~ 120 ℃ Can be.

본 발명에 있어서, 상기 (d) 단계의 알코올은 methanol (C1)을 포함한 1차, 2차, 3차 구조의 모든 알코올인 것을 특징으로 할 수 있다.In the present invention, the alcohol of step (d) may be characterized in that all alcohols of primary, secondary, tertiary structure including methanol (C1).

본 발명의 일 양태에서는 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}생성능을 가지는 Alacligenes latus를 배양하여 폴리-3-하이드록 시부티레이트을 생성시켰으며, 상기 배양은 균체증식을 위한 1단계 배양과 폴리-3-하이드록시부티레이트 생성을 위한 2단계 배양으로 나누어 수행하였다. 상기 폴리-3-하이드록시부티레이트 생성을 위한 2단계 배양은 질소원을 제한한 배지를 사용하여 폴리-3-하이드록시부티레이트의 생성을 유도하였다.In one embodiment of the present invention by culturing Alacligenes latus having a poly-3-hydroxybutyrate (poly (3-hydroxybutyrate)) production to produce a poly-3-hydroxy sibutyrate, the culture is one step for cell growth The dividing was carried out in two stages of culturing and poly-3-hydroxybutyrate production. The two-stage culture for the poly-3-hydroxybutyrate production induced the production of poly-3-hydroxybutyrate using a medium with a limited nitrogen source.

본 발명의 바람직한 양태에서, Alacligenes latus에 의해 생성된 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}은 in vivo 상태에서 배양액의 pH를 4로 조절하고 30분간 정치하여, 자가분해시켜 3-하이드록시부티레이트(폴리-3-하이드록시부티레이트 단량체)을 제조하였다. 상기 배양액의 폴리-3-하이드록시부티레이트의 농도는 0.42g/L이었으며, pH를 조절한 후 정치반응하여 수득된 용액 내의 3-하이드록시부티레이트가 농도는 0.42g/L로 약 95%의 수율로 3-하이드록시부티레이트가 생성되었다.In a preferred embodiment of the present invention, poly-3-hydroxybutyrate produced by Alacligenes latus is adjusted to pH 4 of the culture in vivo and left to stand for 30 minutes, followed by autolysis. -Hydroxybutyrate (poly-3-hydroxybutyrate monomer) was prepared. The concentration of poly-3-hydroxybutyrate in the culture solution was 0.42 g / L, and 3-hydroxybutyrate in the solution obtained by standing reaction after adjusting the pH was 0.42 g / L at a yield of about 95%. 3-hydroxybutyrate was produced.

본 발명의 바람직한 양태에서, 상기 수득된 3-하이드록시부티레이트 용액은 동결건조하여 물을 제거한 후, 클로로포름을 첨가하고, H2SO4를 포함하는 메탄올을 첨가한 후, 100℃에서 12시간 반응시키고, 물을 첨가한 다음, 유기용매층을 분리하여 3-하이드록시부티레이트 메틸에스테르를 수득하였으며, 생성된 3-하이드록시부티레이트 메틸에스테르(R3HB methyl ester)의 농도는 0.40g/L이였으며, 이는 100%의 수율로 3-하이드록시부티레이트로부터 3-하이드록시부티레이트 메틸에스테르로의 전환이 이루어 졌음을 의미한다.In a preferred embodiment of the present invention, the 3-hydroxybutyrate solution obtained is lyophilized to remove water, and then chloroform is added, methanol containing H 2 SO 4 is added, and then reacted at 100 ° C. for 12 hours. After addition of water, the organic solvent layer was separated to give 3-hydroxybutyrate methyl ester, and the concentration of the resulting 3-hydroxybutyrate methyl ester (R3HB methyl ester) was 0.40 g / L, which was 100 Mean conversion of 3-hydroxybutyrate to 3-hydroxybutyrate methylester in% yield.

다른 관점에서, 본 발명은 (a) 폴리하이드록시알케노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알케노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알케노에이트를 함유하는 미생물에서 폴리하이드록시알케노에이트를 자가분해시켜, 하이드록시알케노에이트를 제조하는 단계; 및 (c) 상기 제조된 하이드록시알케노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알케노에이트 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알케노에이트 알킬에스테르의 제조방법에 관한 것이다.In another aspect, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxyalkenoate generating ability, to produce a polyhydroxyalkenoate; (b) self-degrading polyhydroxyalkenoate in the microorganism containing polyhydroxyalkenoate to prepare hydroxyalkenoate; And (c) adding an alcohol to the prepared hydroxyalkenoate and reacting to prepare a hydroxyalkenoate alkylester.

본 발명에 있어서, 하이드록시알케노에이트는 방향족 고리기, 에폭시기, 시아노기, 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것을 사용할 수 있으나, 이에 한정되는 것은 아니다. In the present invention, hydroxyalkenoate may be modified with a substituent selected from the group consisting of an aromatic ring group, an epoxy group, a cyano group, and a halogen group, but is not limited thereto.

또 다른 관점에서, 본 발명은 (a) 폴리하이드록시알키노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알키노에이트를 생성시키는 단계; (b) 상기 폴리하이드록시알키노에이트를 함유하는 미생물에서 폴리하이드록시알키노에이트를 자가분해시켜, 하이드록시알키노에이트를 제조하는 단계; 및 (c) 상기 제조된 하이드록시알키노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알키노에이트 알킬에스테르를 제조하는 단계를 포함하는 하이드록시알키노에이트 알킬에스테르의 제조방법에 관한 것이다.In another aspect, the present invention comprises the steps of (a) culturing a microorganism having a polyhydroxyalkynoate production ability, to produce a polyhydroxyalkynoate; (b) self-degrading polyhydroxyalkynoate in the microorganism containing polyhydroxyalkynoate to prepare hydroxyalkynoate; And (c) adding an alcohol to the prepared hydroxyalkynoate and reacting to prepare a hydroxyalkynoate alkylester.

본 발명에 있어서, 하이드록시알키노에이트는 방향족 고리기, 에폭시기, 시아노기, 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것을 사용할 수 있으나, 이에 한정되는 것은 아니다. In the present invention, hydroxyalkynoate may be modified with a substituent selected from the group consisting of an aromatic ring group, an epoxy group, a cyano group, and a halogen group, but is not limited thereto.

본 발명은 (a) 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성 능을 가지는 미생물을 배양하여, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}를 생성시키는 단계; (b) 상기 생성된 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}을 함유하는 미생물에서 폴리-3-하이드록시부티레이트을 자가분해시켜, 3-하이드록시부티레이트를 제조하는 단계; 및 (c) 상기 3-하이드록시부티레이트에 메탄올을 첨가하고, 반응시켜 3-하이드록시부티레이트 메틸에스테르를 제조하는 단계를 포함하는 3-하이드록시부티레이트 메틸에스테르의 제조방법에 관한 것이다.The present invention comprises the steps of (a) culturing a microorganism having a poly-3-hydroxybutyrate (poly (3-hydroxybutyrate)) producing ability, to produce a poly-3-hydroxybutyrate (poly (3-hydroxybutyrate)}; (b) self-degrading poly-3-hydroxybutyrate in a microorganism containing the produced poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)} to prepare 3-hydroxybutyrate; And (c) adding methanol to the 3-hydroxybutyrate and reacting to prepare 3-hydroxybutyrate methyl ester.

본 발명에 있어서, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성능을 가지는 미생물은 Alacligenes latus인 것을 특징으로 할 수 있고, 상기 (a) 단계의 배양은 질소원이 제한된 배지에서 수행하는 것을 특징으로 할 수 있다. In the present invention, the microorganism having the ability to produce poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)} may be characterized in that the Alacligenes latus , the culture of step (a) is carried out in a medium with a limited nitrogen source It may be characterized by.

본 발명에 있어서, 상기 (d) 단계의 반응은 80~120℃에서 1~24시간 수행하는 것을 특징으로 할 수 있으며, 상기 (e) 단계의 유기용매는 클로로포름인 것을 특징으로 할 수 있다.In the present invention, the reaction of the step (d) may be performed for 1 to 24 hours at 80 ~ 120 ℃, the organic solvent of the (e) step may be characterized in that the chloroform.

이하 본 발명을 실시예에 의하여 더욱 상세하게 설명한다. 이들 실시예는 단지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 국한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited to these examples.

특히, 하기 실시예에서는 대표적인 폴리하이드록시알카노에이트 및 하이드록 시알카노에이트의 일종인 폴리-3-하이드록시부티레이트 {poly(3-hydroxybutyrate)} 중합체, 3-하이드록시부티레이트 (3-hydroxybutyrate) 단량체 및 3-하이드록시부티레이트 메틸에스테르를 생산하였으나, 본 발명에서의 하이드록시알카노에이트는 3번째 위치(3-hydroxy)를 포함하는 여러 가지 다른 위치의 하이드록시기의 알카노에이트 (alkanoate)를 모두 포함하고, 알카노에이트 뿐 아니라, 알케노이트 (alkenoate)와 알키노에이트 (alkynoate) 또한 포함한다는 것 역시 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.In particular, in the following examples, a poly-3-hydroxybutyrate polymer, a 3-hydroxybutyrate monomer, which is a kind of representative polyhydroxyalkanoate and hydroxyalkanoate, And 3-hydroxybutyrate methyl ester, but the hydroxyalkanoate in the present invention is alkanoate of the hydroxy group in various other positions including the third position (3-hydroxy) And alkanoate as well as alkenoate and alkynoate will be apparent to those of ordinary skill in the art.

하기 실시예에서는, 하이드록시알카노에이트는 하이드록시기 뿐 아니라, 방향족 고리기, 에폭시기, 시아노기, 또는 할로겐 원소 등으로 추가로 수식된 것을 포함하는 것 역시 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.In the following examples, hydroxyalkanoate is not only a hydroxy group, but also includes those further modified with an aromatic ring group, an epoxy group, a cyano group, or a halogen element, etc. to those skilled in the art Will be self-evident.

하기 실시예에서는 Alcaligenes latus를 숙주 미생물로 이용하였으나, 다른 대장균이나, 박테리아, 효모 및 곰팡이를 사용하여 poly(3-hydroxybutyrate) 생성능을 향상시키고, 3-hydroxybutyrate 단량체로부터 3-HB alkylester를 제조하는 것 역시 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.In the following examples, Alcaligenes latus was used as the host microorganism, but poly (3-hydroxybutyrate) was used using other E. coli or bacteria, yeast and mold. Improving production capacity and preparing 3-HB alkylesters from 3-hydroxybutyrate monomers will also be apparent to those of ordinary skill in the art.

또한, 하기 실시예에서는 poly(3-hydroxybutyrate)를 이용하여, 3-hydroxybutyrate alkylester를 제조하는 방법만을 기재하였으나, poly(3-hydroxybutyrate)가 속하는 폴리하이드록시알카노에이트를 이용하여, 하이드록시알카노에이트 알킬에스테르를 제조할 수 있다는 것은 당업계에서 통상의 지식을 가진 자에게 있어서 자명할 것이다.In addition, in the following examples, only a method of preparing 3-hydroxybutyrate alkylester using poly (3-hydroxybutyrate) was described, but hydroxyalkano using polyhydroxyalkanoate to which poly (3-hydroxybutyrate) belongs It will be apparent to one of ordinary skill in the art that an alkylene ester can be prepared.

또한, 하기 실시예에서는 3-hydroxybutyrate의 esterification 과정에서 메 탄올(methanol)만을 사용하였으나, 기타 alcohol을 사용하여 3-hydroxybutyrate 및 hydroxyalkanoate를 esterification 시킴으로써 다양한 종류의 3-hydroxybutyrate alkylester 및 hydroxyalkanoate akylester를 제조하는 것 역시 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.       In addition, in the following examples, only methanol was used in the esterification process of 3-hydroxybutyrate, but it is also possible to prepare various types of 3-hydroxybutyrate alkylester and hydroxyalkanoate akylester by esterifying 3-hydroxybutyrate and hydroxyalkanoate using other alcohols. It will be apparent to those of ordinary skill in the art.

아울러, 하기 실시예에서는 특정 배지와 배양방법만을 예시하였으나, 문헌에 보고된 바와 같이, 유청(whey), CSL(corn steep liquor) 등의 당화액과 다른 배지를 사용한 경우나, 유가배양(fed-batch culture), 연속배양 등 다양한 방법을 사용하는 것 (Lee et al., Bioprocess Biosyst. Eng., 26: 63, 2003; Lee et al., Appl. Microbiol. Biotechnol., 58: 663, 2002; Lee et al., Biotechnol. Lett., 25: 111, 2003; Lee et al., Appl. Microbiol. Biotechnol., 54: 23, 2000; Lee et al., Biotechnol. Bioeng., 72: 41, 2001)도 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다. In addition, in the following examples, only a specific medium and a culture method are illustrated, but as reported in the literature, when a saccharified solution such as whey or corn steep liquor (CSL) is used, or when a medium is fed, batch culture), using various methods such as continuous culture (Lee et al. , Bioprocess Biosyst. Eng. , 26: 63, 2003; Lee et al. , Appl. Microbiol. Biotechnol. , 58: 663, 2002; Lee et al. , Biotechnol. Lett. , 25: 111, 2003; Lee et al. , Appl. Microbiol. Biotechnol. , 54: 23, 2000; Lee et al. , Biotechnol. Bioeng. , 72: 41, 2001). It will be apparent to those of ordinary skill in the art.

실시예: Example Alcaligenes latusAlcaligenes latus 균주를 이용한 (R)-3-하이드록시부티레이트 메틸에스테르의 생산  Production of (R) -3-hydroxybutyrate Methyl Ester Using Strains

(1) (One) Alcaligenes latusAlcaligenes latus 균주의 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}생산을 위한 배양  Culture for Production of Poly-3-hydroxybutyrate

Alacligenes latus ATCC 29712 균주를 배양하는데 있어서, 배지 내에 질소원(nitrogen)을 제한하여, poly(3-hydroxybutyrate) {P(3HB)} 생산을 하기 위하여, 2단계 배양을 시행하였다(Wang, F. and Lee, S. Y., Appl. Environ. Microbiol., 63: 3703, 1997; 대한민국 특허등록 제199,995호). In culturing Alacligenes latus ATCC 29712, two-step culture was performed to limit the nitrogen in the medium and produce poly (3-hydroxybutyrate) {P (3HB)} (Wang, F. and Lee). , SY, Appl. Environ.Microbiol ., 63: 3703, 1997; Korean Patent Registration No. 199,995).

먼저, 1 단계배양으로는, A. latus 균주를 100 ml 의 nutrient broth (NB)를 함유한 250 ml 플라스크에서 30 ℃, 250 rpm 으로 24시간 배양하였다. 상기 배양액을 6000rpm에서 10분간 원심분리하여 균체를 회수하고, 2단계 배양에 사용되어지는 AL1 배지로 세포에 남아있는 NB 배지성분을 세척한 후, 다시 6000rpm에서 10분간 원심분리하여 균체를 회수하고, 100ml의 AL1 배지에 현탁시켰다. AL1 배지(pH 7.0)의 조성은 다음과 같다: 증류수 1 liter 당 KH2PO4 1.5 g, Na2HPO4ㅇ12H2O 9 g, MgSO4ㅇ7H2O 0.2 g, CaCl2ㅇ2H2O 0.01 g, citric acid 0.1 g, trace element solution 1 ml. First, as a one-step culture, A. latus strains were incubated at 250C for 24 hours at 250C in a 250 ml flask containing 100 ml of nutrient broth (NB). The cells were recovered by centrifuging the culture solution at 6000 rpm for 10 minutes, washing the NB medium components remaining in the cells with AL1 medium used for the two-step culture, and then recovering the cells by centrifuging at 6000 rpm for 10 minutes. It was suspended in 100 ml of AL1 medium. The composition of the AL1 medium (pH 7.0) is as follows: Distilled water 1 KH 2 PO 4 1.5 per liter g, Na 2 HPO 4 o 12H 2 O 9 g, MgSO 4 o 7H 2 O 0.2 g, CaCl 2 o 2H 2 O 0.01 g, citric acid 0.1 g, trace element solution 1 ml.

Trace element solution의 조성은 다음과 같다: 증류수 1 liter 당 FeSO4ㅇ7H2O 20 g, H3BO4 0.3 g, CoCl2ㅇ6H2O 0.2 g, ZnSO4ㅇ7H2O 0.03 g, MnCl2ㅇ4H2O 0.03 g, (NH4)6Mo7O24ㅇ4H2O 0.03 g, NiSO4ㅇ7H2O 0.03 g, CuSO4ㅇ5H2O 0.01 g . The composition of the trace element solution is as follows: 20 g of FeSO 4 ㅇ 7H 2 O, 1 g of H 3 BO 4 , CoCl 2 ㅇ 6H 2 O 0.2 g, ZnSO 4 ㅇ 7H 2 O 0.03 g, MnCl 2 per liter of distilled water 0.03 g of 4H 2 O, (NH 4 ) 6 Mo 7 O 24 0.03 g of 4H 2 O, 0.03 g of NiSO 4 7H 2 O, 0.01 g of CuSO 4 −5H 2 O.

100ml의 AL1 배지에 현탁시킨 세균은 탄소원으로 20 g/l의 수크로오스를 첨가한 후, 30 ℃, 250 rpm 으로 24시간 배양하였다.. 배양 종료 후, 배양액을 6000 rpm,에서 10분간 원심분리하여 세포를 회수하였다. 회수한 세포를 증류수로 한 번 씻어낸 후 100 ℃의 건조기에서 24시간 건조하였다. Bacteria suspended in 100 ml of AL1 medium were incubated for 24 hours at 30 ° C. and 250 rpm after adding 20 g / l sucrose as a carbon source. After completion of the culture, the culture medium was centrifuged at 6000 rpm for 10 minutes for cells. Was recovered. The collected cells were washed once with distilled water and dried in a drier at 100 ° C. for 24 hours.

건조된 세포로 가스크로마토그래피(gas chromatography) 분석을 수행하여, 세포 내 합성된 P(3HB) 함량을 측정하였다. 2 단계 플라스크 배양 결과, 총 건조세포무게(농도)는 0.48 g/l 이었고, 그 중 P(3HB) 무게(농도)는 0.42 g/l 로, 세포 내에 약 88 wt%의 P(3HB)가 생성된 것을 확인하였다.Gas chromatography analysis was performed on the dried cells, and the synthesized P (3HB) content in the cells was measured. As a result of the two-step flask culture, the total dry cell weight (concentration) was 0.48 g / l, of which P (3HB) weight (concentration) was 0.42 g / l, which produced about 88 wt% of P (3HB) in the cells. It was confirmed.

(2) (2) in vivoin vivo 에서의In 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}의 depolymerization을 통한 (R)-3-하이드록시부티레이트의 생산 Production of (R) -3-hydroxybutyrate through depolymerization of poly-3-hydroxybutyrate

(1)의 2 단계 배양이 끝난 A. latus 배양액에 H2SO4를 첨가하여 배양액의 pH를 4가 되도록 조정한 후, 37 ℃에서 30분 동안 정치(no shaking)하였다. 그 후, 배양액을 6000 rpm에서 10분간 원심분리하여, 상등액(supernatant)을 분리하였다. 상등액에 녹아있는 (R)-3-hydroxybutyrate (R3HB) 의 농도를 측정하기 위해 고성능 리퀴드크로마토그래피(high-performance liquid chromatography) 분석을 수행하였다. 그 결과, 상등액의 R3HB 농도는 0.40 g/l 로, 약 95%의 수율로 P(3HB)로부터 생성되었다 (Lee, S.Y. et al., Biotechnol. Bioeng., 65:363, 1999; 한국등록특허 제 250,830호).After (1) a two-step culture A. ended by the addition of H 2 SO 4 in latus culture medium adjusted to 4. The pH of the culture solution, the mixture was allowed to stand (no shaking) for 30 minutes at 37 ℃. The culture was then centrifuged at 6000 rpm for 10 minutes to separate the supernatants. High-performance liquid chromatography analysis was performed to determine the concentration of (R) -3-hydroxybutyrate (R3HB) dissolved in the supernatant. As a result, the R3HB concentration of the supernatant was 0.40 g / l, which was produced from P (3HB) in a yield of about 95% (Lee, SY et al., Biotechnol. Bioeng. , 65: 363, 1999; 250,830).

(3) (R)-3-hydroxybutyrate (R3HB) 에서 R3HB methyl ester 로의 전환(3) Conversion of (R) -3-hydroxybutyrate (R3HB) to R3HB methyl ester

상기 (2)에서 얻은 상등액에 녹아있는 R3HB 형태를 동결건조를 통해 물을 제거하였다. 물이 제거된 R3HB에 클로로포름 2ml을 첨가하고, 3% (v/v) H2SO4 를 포함하는 메탄올 1 ml 을 첨가한 후, 이 혼합물을 100 ℃에서 12 시간 동안 반응시켰 다. 반응이 끝난 후, 혼합물을 상온까지 식히고, 혼합물에 증류수 1ml을 첨가하여 5분 동안 격렬히 섞어 주어, 유기용매(chloroform) 층과 물(수용액) 층이 분리시키고, 10,000 rpm에서 10분간 원심분리하여, 유기용매 층만 채취하여 가스크로마토그래피(gas chromatography) 분석을 수행함으로써, 생성된 R3HB 메틸 에스테르 농도를 측정하였다. 그 결과, R3HB methyl ester 농도는 0.40g/l로, 100%의 수율로 R3HB로부터 전환되었다. 클로로포름에 녹아있는 R3HB methyl ester 형태에서 용매인 클로로포름을 증발시켜 R3HB 메틸에스테르를 얻었다. R3HB forms dissolved in the supernatant obtained in (2) were removed by lyophilization. 2 ml of chloroform was added to R 3 HB from which water was removed, and 1 ml of methanol containing 3% (v / v) H 2 SO 4 was added, and the mixture was reacted at 100 ° C. for 12 hours. After the reaction was completed, the mixture was cooled to room temperature, 1 ml of distilled water was added to the mixture and mixed vigorously for 5 minutes. The organic solvent (chloroform) layer and the water (aqueous) layer were separated, and centrifuged at 10,000 rpm for 10 minutes. Only the organic solvent layer was taken, and then subjected to gas chromatography, thereby measuring the concentration of the produced R3HB methyl ester. As a result, the R3HB methyl ester concentration was 0.40 g / l, converted from R3HB in 100% yield. R3HB methyl ester was obtained by evaporating the solvent chloroform in the form of R3HB methyl ester dissolved in chloroform.

비교예: poly(3-hydroxybutyrate)을 이용한 R3HB methyl ester의 in vitro생산Comparative Example: In vitro Production of R3HB Methyl Ester Using Poly (3-hydroxybutyrate)

시판되는 P(3HB)(Jiangsu, China)를 사용하여, in vitro에서 R3HB methyl ester를 제조하였다 (Xiaojun Zhang et al., Biomacromolecules, 10: 707, 2009).R3HB methyl ester was prepared in vitro using commercially available P (3HB) (Jiangsu, China). (Xiaojun Zhang et al ., Biomacromolecules , 10: 707, 2009).

15 g의 PHB를 200mL의 클로로포름용액에 용해시키고, 200mL의 산성메탄올(15%(v/v) H2SO4 in methanol)을 첨가하고, 100℃에서 1시간 환류시켰다. 상기 혼합액을 실온에서 냉각시킨 다음, 40mL의 포화 NaCl용액을 첨가하고, 10분간 교반하여, 유기용매층과 물층을 분리하였다. 물층을 50mL의 클로로포름으로 3회 추출하고, 건조시킨 후, Na2SO4를 이용하여, 유기용매층과 혼합하고, 진공상태에서 증발시켜, 점성있는 투명한 액체인 R3HB methyl ester를 수득하였다. 15 g of PHB was dissolved in 200 mL of chloroform solution, 200 mL of acid methanol (15% (v / v) H 2 SO 4 in methanol) was added, and the mixture was refluxed at 100 ° C for 1 hour. After cooling the mixture at room temperature, 40 mL of saturated NaCl solution was added and stirred for 10 minutes to separate the organic solvent layer and the water layer. The water layer was extracted three times with 50 mL of chloroform, dried, and then mixed with an organic solvent layer using Na 2 SO 4 , and evaporated in vacuo to give a viscous transparent liquid, R 3 HB methyl ester.

비교예에서 수득한 R3HB methyl ester와 실시예에서 수득한 R3HB methyl ester의 전환율을 측정한 결과, P(3HB)를 in vitro 전환시킨 실시예의 R3HB methyl ester 최종수율은 95%로, 비교예의 최종수율 52%보다 월등히 높은 생산수율을 나타내었다(표 1).As a result of measuring the conversion rate of the R3HB methyl ester obtained in the Comparative Example and the R3HB methyl ester obtained in the Example, the final yield of the R3HB methyl ester of the Example in which P (3HB) was converted in vitro was 95%. The production yield was much higher than% (Table 1).

실시예와 비교예의 R3HB methyl ester 전환율 비교 Comparison of R3HB methyl ester conversion rate in Example and Comparative Example 비교예Comparative example 실시예Example
시작물질
Starting material
시판 P(3HB)
[정제된 P(3HB)]
Commercially available P (3HB)
Purified P (3HB) 미생물균주배양을 통한
P(3HB) 직접 생산
[미생물세포내에 축적된 P(3HB)]Through microbial strain culture
P (3HB) Direct Production
[P (3HB) Accumulated in Microbial Cells]


전환방법


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산을 촉매로한 P(3HB)의 가수분해

Hydrolysis of P (3HB) by Acid Catalyst
세포내에서 P(3HB)의 생분해를 통한 R3HB 생산 [전환수율: 95%]R3HB production through biodegradation of P (3HB) intracellularly [Conversion yield: 95%] 산을 촉매로 한 에스테르화 반응을 통한 R3HB로부터
R3HB methyl ester의 생산
[전환수율: 100%]From R3HB through acid-catalyzed esterification
Production of R3HB Methyl Ester
[Conversion yield: 100%] 최종산물Final product R3HB methyl esterR3HB methyl ester R3HB methyl esterR3HB methyl ester 최종수율Final yield 52%52% 95%95%

이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적 기술은 단지 바람직한 실시태양일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.  The specific parts of the present invention have been described in detail above, and it is apparent to those skilled in the art that such specific descriptions are merely preferred embodiments, and thus the scope of the present invention is not limited thereto. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

도 1은 A. latus ATCC 29712 균주를 이용하여 3HB methyl ester를 고효율로 제조하는 공정을 나타낸 것이다.Figure 1 shows a process for producing 3HB methyl ester with high efficiency using A. latus ATCC 29712 strain.

Claims (20)

다음 단계를 포함하는 하이드록시알카노에이트 알킬에스테르의 제조방법:Method for preparing a hydroxyalkanoate alkyl ester comprising the following steps: (a) 폴리하이드록시알카노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알카노에이트를 생성시키는 단계;(a) culturing a microorganism having a polyhydroxyalkanoate generating ability to produce polyhydroxyalkanoate; (b) 상기 폴리하이드록시알카노에이트를 함유하는 미생물에서 폴리하이드록시알카노에이트를 자가분해시켜, 하이드록시알카노에이트를 제조하는 단계; 및(b) self-degrading polyhydroxyalkanoate in the microorganism containing polyhydroxyalkanoate to prepare hydroxyalkanoate; And (c) 상기 제조된 하이드록시알카노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알카노에이트 알킬에스테르를 제조하는 단계.(c) adding an alcohol to the prepared hydroxyalkanoate and reacting to prepare a hydroxyalkanoate alkyl ester. 제1항에 있어서, 하이드록시알카노에이트는 방향족 고리기, 에폭시기, 시아노기 및 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것임을 특징으로 하는 방법.The method of claim 1, wherein the hydroxyalkanoate is modified with a substituent selected from the group consisting of aromatic ring groups, epoxy groups, cyano groups and halogen groups. 제1항에 있어서, (b) 단계의 자가분해는 폴리하이드록시알카노에이트를 함유하는 미생물을 함유하는 배양액의 pH를 2~5로 조절하고, 34~50℃에서 정치반응시켜 수행하는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.According to claim 1, wherein the self-decomposition of step (b) is carried out by adjusting the pH of the culture medium containing the microhydroxyalkanoate containing 2 to 5, and the reaction at 34 ~ 50 ℃ The manufacturing method of the hydroxy alkanoate alkyl ester which is used. 제1항에 있어서, (c) 단계는 유기용매를 추가로 첨가하여 수행하는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.The method for preparing hydroxyalkanoate alkyl ester according to claim 1, wherein step (c) is performed by further adding an organic solvent. 제1항에 있어서, 폴리하이드록시알카노에이트 생성능을 가지는 미생물은 Aeromonas 속 미생물, Achromobacter 속 미생물, Acidovorax delafieldii, Acidovax facilis,, Acinetobacter 속 미생물, Actinomyces 속 미생물, Aeromonas 속 미생물, Alcaligenes 속 미생물, Alteromonas 속 미생물, Amoebobacter 속 미생물, Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp., Azospirillum 속 미생물, Azotobacter 속 미생물, Bacillus 속 미생물, Beggiatoa 속 미생물, Beijerinckia 속 미생물, Beneckea 속 미생물, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter 속 미생물, Chlorogloea 속 미생물, Chromatium 속 미생물, Chromobacterium 속 미생물, Clostridium 속 미생물, Comamonas 속 미생물, Corynebacterium 속 미생물, Cyanobacteria 속 미생물, Derxia 속 미생물, Desulfonema 속 미생물, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira 속 미생물, Ferrobacillus ferroxidans, Flavobacterium sp., Haemophilus influenzae, Halobacterium 속 미생물, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, Hydrogenophaga 속 미생물, Hyphomicrobium 속 미생물, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., Leptothrix discophorus, Methylobacterium 속 미생물, Methylosinus 속 미생물, Micrococcus 속 미생물, Mycobacterium 속 미생물, Nitrobacter 속 미생물, Nocardia 속 미생물, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium 속 미생물, Physarum ploycephalum, Pseudomonas 속 미생물, Ralstonia 속 미생물, Rhizobium 속 미생물, Rhodobacillus 속 미생물, Rhodobacter 속 미생물, Rhodococcus 속 미생물, Rhodocyclus 속 미생물, Rhodomicrobium vannielii, Rhodopseudomonas 속 미생물, Rhodospirillum 속 미생물, Sphingomonas paucimobilis, Spirillum 속 미생물, Spirulina 속 미생물, Staphylococcus 속 미생물, Stella 속 미생물, Streptomyces 속 미생물, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus 속 미생물, Thiocapsa 속 미생물, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea 속 및 폴리하이드록시알카노에이트 생성능을 가지는 효소를 코딩하는 유전자로 형질전환된 미생물로 구성된 군에서 선택되는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.The microorganism having a polyhydroxyalkanoate- producing ability according to claim 1, wherein the microorganism having a polyhydroxyalkanoate-generating ability is a genus Aeromonas , Achromobacter genus, Acidovorax delafieldii , Acidovax facilis ,, Acinetobacter genus, Actinomyces microorganism, Aeromonas genus, Alcaligenes genus, Alteromonas genus Microorganism, Amoebobacter spp ., Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp ., Azospirillum spp, Azotobacter spp, Bacillus spp, Beggiatoa spp, Beijerinckia spp, Beneckea spp, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter spp, Chlorogloea spp ., Chromatium microorganism of the genus, Chromobacterium spp, Clostridium spp, Comamonas spp, Corynebacterium spp, Cyanobacteria in microorganisms, Derxia spp, Desulfonema spp, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira spp, Ferrobacillus ferroxidans, Flavobacterium sp, Haemophilus influenzae, Halobacterium spp, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, microorganisms in Hydrogenophaga, microorganisms in Hyphomicrobium, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., L eptothrix discophorus, Methylobacterium spp, Methylosinus spp, Micrococcus spp, Mycobacterium spp, Nitrobacter spp, Nocardia spp, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium spp, Physarum ploycephalum, Pseudomonas spp, Ralstonia spp , Rhizobium spp, Rhodobacillus spp, Rhodobacter spp, Rhodococcus spp, Rhodocyclus spp, Rhodomicrobium vannielii, Rhodopseudomonas spp, Rhodospirillum spp, Sphingomonas paucimobilis, Spirillum spp, Spirulina spp, Staphylococcus spp, Stella spp , Streptomyces spp, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus spp, Thiocapsa spp, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea in And a microorganism transformed with a gene encoding an enzyme having a polyhydroxyalkanoate generating ability. 제1항에 있어서, 하이드록시알카노에이트는 락테이트(lactate), 2-하이드록시부탄산(hydroxybutyrate), 3-하이드록시프로피온산(hydroxypropionate), 3-하이드록시부티레이트(hydroxybutyrate), 3-하이드록시발레르산(hydroxyvalerate), 4-하이드록시부탄산(hydroxybutyrate), 탄소수가 6~14개인 중간사슬 길이의 (D)-3-하이드록시카르복실산(hydroxycarboxylic acids), 3-하이드록시프로피온산(hydroxypropionic acid), 3-하이드록시헥산산(hydroxyhexanoic acid), 3-하이드록시헵탄산(hydroxyheptanoic acid), 3-하이드록시옥탄산(hydroxyoctanoic acid), 3-하이드록시노난산(hydroxynonanoic acid), 3-하이드록시데칸산(hydroxydecanoic acid), 3-하이드록시운데칸산(hydroxyundecanoic acid), 3-하이드록시 도데칸산(hydroxydodecanoic acid), 3-하이드록시테트라데칸산(hydroxytetradecanoic acid), 3-하이드록시헥사데칸산(hydroxyhexadecanoic acid), 4-하이드록시발레르산(hydroxyvaleric acid), 4-하이드록시헥산산(hydroxyhexanoic acid), 4-하이드록시헵탄산(hydroxyheptanoic acid), 4-하이드록시옥탄산(hydroxyoctanoic acid), 4-하이드록시데칸산(hydroxydecanoic acid), 5-하이드록시발레르산(hydroxyvaleric acid), 5-하이드록시헥산산(hydroxyhexanoic acid), 6-하이드록시도데칸산(hydroxydodecanoic acid), 3-하이드록시(hydroxy)-4-펜텐산(pentenoic acid), 3-하이드록시(hydroxy)-4-trans-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-4-cis-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-5-헥센산(hexenoic acid), 3-하이드록시(hydroxy)-6-trans-옥텐산(octenoic acid), 3-하이드록시(hydroxy)-6-cis-옥텐산(octenoic acid), 3-하이드록시(hydroxy)-7-옥텐산(octenoic acid), 3- 하이드록시(hydroxy)-8-노넨산(nonenoic acid), 3-하이드록시(hydroxy)-9-데센산(decenoic acid), 3-하이드록시(hydroxy)-5-cis-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-6-cis-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-5-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-7-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-5,8-cis-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-4-메틸발레르산(methylvaleric acid), 3-하이드록시(hydroxy)-4-메틸헥산산(methylhexanoic acid), 3-하이드록시(hydroxy)-5-메틸헥산산(methylhexanoic acid), 3-하이드록시(hydroxy)-6-메틸헵탄산(methylheptanoic acid), 3-하이드록시(hydroxy)-4-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-5-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-6-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-7-메틸옥탄산(methyloctanoic acid), 3-하이드록시(hydroxy)-6-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-7-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-8-메틸노난산(methylnonanoic acid), 3-하이드록시(hydroxy)-7-메틸데칸산(methyldecanoic acid), 3-하이드록시(hydroxy)-9-메틸데칸산(methyldecanoic acid), 3-하이드록시(hydroxy)-7-메틸-6-옥텐산(octenoic acid), 말산(malic acid), 3-하이드록시숙신산(hydroxysuccinic acid)-메틸에스테르, 3-하이드록시아디핀산(hydroxyadipinic acid)-메틸에스테르, 3-하이드록시스베린산(hydroxysuberic acid)-메틸에스테르, 3-하이드록시아젤라인산(hydroxyazelaic acid)-메틸에스테르, 3-하이드록시세바신산(hydroxysebacic acid,)-메틸에스테르, 3-하이드록시스베린산(hydroxysuberic acid)-에틸에스테르, 3-하이드록시세바신산(hydroxysebacic acid)-에틸에스테르, 3-하이드록시피메린산(hydroxypimelic acid)-프로필에스테르, 3-하이드록시세바신산(hydroxysebacic acid)-벤질에스테르, 3-하이드록시(hydroxy)-8-아세톡시옥탄산(acetoxyoctanoic acid), 3-하이드록시(hydroxy)-9-아세톡시노난산(acetoxynonanoic acid), 페녹시(phenoxy)-3-하이드록시부티레이트(hydroxybutyric acid), 페녹시(phenoxy)-3-하이드록시발레르산(hydroxyvaleric acid), 페녹시(phenoxy)-3-하이드록시헵탄산(hydroxyheptanoic acid), 페녹시(phenoxy)-3-하이드록시옥탄산(hydroxyoctanoic acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시부티레이트(hydroxybutyric acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시발레르산(hydroxyvaleric acid), para-시아노페녹시(cyanophenoxy)-3-하이드록시헥산산(hydroxyhexanoic acid), para-니트로페녹시(nitrophenoxy)-3-하이드록시헥산산(hydroxyhexanoic acid), 3-하이드록시(hydroxy)-5-페닐발레르산(phenylvaleric acid), 3-하이드록시(hydroxy)-6-페닐헥산(phenylhexanoic acid), 3-하이드록시(hydroxy)-7-페닐헵탄산(phenylheptanoic acid), 3-하이드록시(hydroxy)-8-페닐옥탄산(phenyloctanoic acid), 3-하이드록시(hydroxy)-9-페닐노난산(phenylnonanoic acid), 3-하이드록시(hydroxy)-10-페닐데칸산(phenyldecanoic acid), 3-하이드록시(hydroxy)-5-시클로헥실부탄산(cyclohexylbutyric acid), 3,12-디하이드록시도데칸산(dihydroxydodecanoic acid), 3,8-디하이드록시(dihydroxy)-5-cis-테트라데센산(tetradecenoic acid), 3-하이드록시(hydroxy)-4,5-에폭시데칸산(epoxydecanoic acid), 3-하이드록 시(hydroxy)-6,7-에폭시도데칸산(epoxydodecanoic acid), 3-하이드록시(hydroxy)-8,9-에폭시(epoxy)-5,6-cis-테트라데칸산(tetradecanoic acid), 7-시아노(cyano)-3-하이드록시헵탄산(hydroxyheptanoic acid), 9-시아노(cyano)-3-하이드록시노난산(hydroxynonanoic acid), 3-하이드록시(hydroxy)-7-플루오로헵탄산(fluoroheptanoic acid), 3-하이드록시(hydroxy)-9-플루오로노난산(fluorononanoic acid), 3-하이드록시(hydroxy)-6-클로로헥산산(chlorohexanoic acid), 3-하이드록시(hydroxy)-8-클로로옥탄산(chlorooctanoic acid), 3-하이드록시(hydroxy)-6-브로모헥산산(bromohexanoic acid), 3-하이드록시(hydroxy)-8-브로모옥탄산(bromooctanoic acid), 3-하이드록시(hydroxy)-11-브로모운데칸산(bromoundecanoic acid), 3-하이드록시(hydroxy)-2-부텐산(butenoic acid), 6-하이드록시(hydroxy)-3-도데센산(dodecenoic acid), 3-하이드록시(hydroxy)-2-메틸부탄산(methylbutyric acid), 3-하이드록시(hydroxy)-2-메틸발레르산(methylvaleric acid) 및 3-하이드록시(hydroxy)-2,6-디메틸-5-헵텐산(heptenoic acid)로 구성된 군에서 선택된 하나 이상인 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.The method of claim 1, wherein the hydroxyalkanoate is lactate, 2-hydroxybutyrate, 3-hydroxypropionate, 3-hydroxybutyrate, 3-hydroxy Valeric acid, 4-hydroxybutyrate, medium chain length (D) -3-hydroxycarboxylic acids having 6 to 14 carbon atoms, 3-hydroxypropionic acid ), 3-hydroxyhexanoic acid, 3-hydroxyheptanoic acid, 3-hydroxyoctanoic acid, 3-hydroxynonanoic acid, 3-hydroxy Decanoic acid, 3-hydroxyundecanoic acid, 3-hydroxy dodecanoic acid, 3-hydroxytetradecanoic acid, 3-hydroxyhexadecanoic acid acid), 4-hydroxyballet Hydroxyvaleric acid, 4-hydroxyhexanoic acid, 4-hydroxyheptanoic acid, 4-hydroxyoctanoic acid, 4-hydroxydecanoic acid, 5-hydroxyvaleric acid, 5-hydroxyhexanoic acid, 6-hydroxydodecanoic acid, 3-hydroxy-4-pentenoic acid , 3-hydroxy-4-trans-hexenoic acid, 3-hydroxy-4-cis-hexenoic acid, 3-hydroxy-5-hex Hexanoic acid, 3-hydroxy-6-trans-octenic acid, 3-hydroxy-6-cis-octennoic acid, 3-hydroxy ) -7-octenic acid, 3-hydroxy-8-nonenoic acid, 3-hydroxy-9-decenoic acid, 3-hydroxy ( hydroxy-5-cis-dodecenoic acid, 3-hydroxy-6-cis Dodecenoic acid, 3-hydroxy-5-cis-tetradecenoic acid, 3-hydroxy-7-cis-tetradecenoic acid, 3- Hydroxy-5,8-cis-cis-tetradecenoic acid, 3-hydroxy-4-methylvaleric acid, 3-hydroxy-4- Methylhexanoic acid, 3-hydroxy-5-methylhexanoic acid, 3-hydroxy-6-methylheptanoic acid, 3-hydroxy ) -4-methyloctanoic acid, 3-hydroxy-5-methyloctanoic acid, 3-hydroxy-6-methyloctanoic acid, 3- Hydroxy-7-methyloctanoic acid, 3-hydroxy-6-methylnonanoic acid, 3-hydroxy-7-methylnonanoic acid ), 3-hydroxy-8-methylnonanoic acid, 3- Hydroxy-7-methyldecanoic acid, 3-hydroxy-9-methyldecanoic acid, 3-hydroxy-7-methyl-6-octenic acid (octenoic acid), malic acid, 3-hydroxysuccinic acid-methyl ester, 3-hydroxyadipinic acid-methyl ester, 3-hydroxysuberic acid- Methyl ester, 3-hydroxyazelaic acid-methyl ester, 3-hydroxysebacic acid-methyl ester, 3-hydroxysuberic acid-ethyl ester, 3-hydroxy Hydroxysebacic acid-ethyl ester, 3-hydroxypimelic acid-propyl ester, 3-hydroxysebacic acid-benzyl ester, 3-hydroxy-8- Acetoxyoctanoic acid, 3-hydroxy-9-acetoxynonanoi c acid), phenoxy-3-hydroxybutyric acid, phenoxy-3-hydroxyvaleric acid, phenoxy-3-hydroxyheptanoic acid ( hydroxyheptanoic acid, phenoxy-3-hydroxyoctanoic acid, para-cyanophenoxy-3-hydroxybutyric acid, para-cyanophenoxy -3-hydroxyvaleric acid, para-cyanophenoxy-3-hydroxyhexanoic acid, para-nitrophenoxy-3-hydroxyhexanoic acid), 3-hydroxy-5-phenylvaleric acid, 3-hydroxy-6-phenylhexane, 3-hydroxy-7-phenylheptane Phenylheptanoic acid, 3-hydroxy-8-phenyloctanoic acid, 3-hydroxy-9-phenylnonanoic acid, 3-hydroxy Hydroxy-10-phenyldecanoic acid, 3-hydroxy-5-cyclohexylbutyric acid, 3,12-dihydroxydodecanoic acid, 3 , 8-dihydroxy-5-cis-tetradecenoic acid, 3-hydroxy-4,5-epoxydecanoic acid, 3-hydroxy -6,7-epoxydodecanoic acid, 3-hydroxy-8,9-epoxy-5,6-cis-tetradecanoic acid, 7-cyano ( cyano-3-hydroxyheptanoic acid, 9-cyano-3-hydroxynonanoic acid, 3-hydroxy-7-fluoroheptanoic acid ), 3-hydroxy-9-fluorononanoic acid, 3-hydroxy-6-chlorohexanoic acid, 3-hydroxy-8-chloro Chlorooctanoic acid, 3-hydroxy-6-bromohexanoic acid hexanoic acid, 3-hydroxy-8-bromooctanoic acid, 3-hydroxy-11-bromooundecanoic acid, 3-hydroxy-2 Butenoic acid, 6-hydroxy-3-dodecenoic acid, 3-hydroxy-2-methylbutyric acid, 3-hydroxy Hydroxyalkanoate alkyl, characterized in that it is at least one selected from the group consisting of 2-methylvaleric acid and 3-hydroxy-2,6-dimethyl-5-heptenoic acid. Process for the preparation of esters. 제1항에 있어서, (a) 단계의 배양은 질소원이 제한된 배지에서 수행하는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.The method of claim 1, wherein the culturing of step (a) is carried out in a medium with a limited nitrogen source. 제1항에 있어서, (c) 단계의 반응은 80~120℃에서 1~24시간 수행하는 것을 특징으로 하는 하이드록시알카노에이트 알킬에스테르의 제조방법.The method of claim 1, wherein the reaction of step (c) is performed at 80 to 120 ° C. for 1 to 24 hours. 다음 단계를 포함하는 하이드록시알케노에이트 알킬에스테르의 제조방법:Method for preparing a hydroxyalkenoate alkyl ester comprising the following steps: (a) 폴리하이드록시알케노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알케노에이트를 생성시키는 단계;(a) culturing a microorganism having a polyhydroxyalkenoate generating ability to produce polyhydroxyalkenoate; (b) 상기 폴리하이드록시알케노에이트를 함유하는 미생물에서 폴리하이드록시알케노에이트를 자가분해시켜, 하이드록시알케노에이트를 제조하는 단계; 및(b) self-degrading polyhydroxyalkenoate in the microorganism containing polyhydroxyalkenoate to prepare hydroxyalkenoate; And (c) 상기 제조된 하이드록시알케노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알케노에이트 알킬에스테르를 제조하는 단계.(c) adding an alcohol to the prepared hydroxyalkenoate and reacting to prepare a hydroxyalkenoate alkyl ester. 제9항에 있어서, 하이드록시알케노에이트는 방향족 고리기, 에폭시기, 시아노기 및 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것임을 특징으로 하는 방법.10. The method of claim 9, wherein the hydroxyalkenoate is modified with a substituent selected from the group consisting of aromatic ring groups, epoxy groups, cyano groups and halogen groups. 다음 단계를 포함하는 하이드록시알키노에이트 알킬에스테르의 제조방법:Method for preparing a hydroxyalkynoate alkyl ester comprising the following steps: (a) 폴리하이드록시알키노에이트 생성능을 가지는 미생물을 배양하여, 폴리하이드록시알키노에이트를 생성시키는 단계;(a) culturing a microorganism having a polyhydroxyalkynoate generating ability to produce polyhydroxyalkynoate; (b) 상기 폴리하이드록시알키노에이트를 함유하는 미생물에서 폴리하이드록시알키노에이트를 자가분해시켜, 하이드록시알키노에이트를 제조하는 단계; 및(b) self-degrading polyhydroxyalkynoate in the microorganism containing polyhydroxyalkynoate to prepare hydroxyalkynoate; And (c) 상기 제조된 하이드록시알키노에이트에 알코올을 첨가하고, 반응시켜 하이드록시알키노에이트 알킬에스테르를 제조하는 단계.(c) adding an alcohol to the prepared hydroxyalkynoate and reacting to prepare a hydroxyalkynoate alkyl ester. 제11항에 있어서, 하이드록시알키노에이트는 방향족 고리기, 에폭시기, 시아노기 및 할로겐기로 구성된 군에서 선택되는 치환기로 수식된 것임을 특징으로 하는 방법.12. The method of claim 11, wherein the hydroxyalkynoate is modified with a substituent selected from the group consisting of aromatic ring groups, epoxy groups, cyano groups and halogen groups. 다음 단계를 포함하는 3-하이드록시부티레이트 메틸에스테르의 제조방법:Process for preparing 3-hydroxybutyrate methyl ester comprising the following steps: (a) 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성능을 가지는 미생물을 배양하여, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}를 생성시키는 단계;(a) culturing a microorganism having a poly-3-hydroxybutyrate} producing ability to produce poly-3-hydroxybutyrate}; (b) 상기 생성된 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}을 함유하는 미생물에서 폴리-3-하이드록시부티레이트을 자가분해시켜, 3-하이드록시부티레이트를 제조하는 단계; 및(b) self-degrading poly-3-hydroxybutyrate in a microorganism containing the produced poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)} to prepare 3-hydroxybutyrate; And (c) 상기 3-하이드록시부티레이트에 메탄올을 첨가하고, 반응시켜 3-하이드록시부티레이트 메틸에스테르를 제조하는 단계.(c) adding methanol to the 3-hydroxybutyrate and reacting to prepare 3-hydroxybutyrate methyl ester. 제13항에 있어서, (b) 단계의 자가분해는 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)}을 함유하는 미생물을 함유하는 배양액의 pH를 2~5로 조절하고, 34~50℃에서 정치반응시켜 수행하는 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The method according to claim 13, wherein the autolysis of step (b) adjusts the pH of the culture medium containing the microorganism containing poly-3-hydroxybutyrate to 2-5, 34-50 Process for the preparation of 3-hydroxybutyrate methyl ester, characterized in that carried out by static reaction at ℃. 제13항에 있어서, (c) 단계는 유기용매를 추가로 첨가하여 수행하는 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The method for preparing 3-hydroxybutyrate methyl ester according to claim 13, wherein step (c) is performed by further adding an organic solvent. 제13항에 있어서, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성능을 가지는 미생물은 Aeromonas 속 미생물, Achromobacter 속 미생물, Acidovorax delafieldii, Acidovax facilis,, Acinetobacter 속 미생물, Actinomyces 속 미생물, Aeromonas 속 미생물, Alcaligenes 속 미생물, Alteromonas 속 미생물, Amoebobacter 속 미생물, Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp., Azospirillum 속 미생물, Azotobacter 속 미생물, Bacillus 속 미생물, Beggiatoa 속 미생물, Beijerinckia 속 미생물, Beneckea 속 미생물, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter 속 미생물, Chlorogloea 속 미생물, Chromatium 속 미생물, Chromobacterium 속 미생물, Clostridium 속 미생물, Comamonas 속 미생물, Corynebacterium 속 미생물, Cyanobacteria 속 미생물, Derxia 속 미생물, Desulfonema 속 미생물, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira 속 미생물, Ferrobacillus ferroxidans, Flavobacterium sp., Haemophilus influenzae, Halobacterium 속 미생물, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, Hydrogenophaga 속 미생물, Hyphomicrobium 속 미생물, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., Leptothrix discophorus, Methylobacterium 속 미생물, Methylosinus 속 미생물, Micrococcus 속 미생물, Mycobacterium 속 미생물, Nitrobacter 속 미생물, Nocardia 속 미생물, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium 속 미생물, Physarum ploycephalum, Pseudomonas 속 미생물, Ralstonia 속 미생물, Rhizobium 속 미생물, Rhodobacillus 속 미생물, Rhodobacter 속 미생물, Rhodococcus 속 미 생물, Rhodocyclus 속 미생물, Rhodomicrobium vannielii, Rhodopseudomonas 속 미생물, Rhodospirillum 속 미생물, Sphingomonas paucimobilis, Spirillum 속 미생물, Spirulina 속 미생물, Staphylococcus 속 미생물, Stella 속 미생물, Streptomyces 속 미생물, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus 속 미생물, Thiocapsa 속 미생물, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea 속 및 폴리-3-하이드록시부티레이트 생성능을 가지는 효소를 코딩하는 유전자로 형질전환된 미생물로 구성된 군에서 선택되는 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The method of claim 13, wherein the microorganism having the ability to produce poly-3-hydroxybutyrate {poly (3-hydroxybutyrate)} is a microorganism of the genus Aeromonas , microorganisms of the genus Achromobacter , Acidovorax delafieldii , Acidovax facilis , microorganisms of the genus Acinetobacter , microorganisms of the genus Actinomyces , Aeromonas Genus microorganism, Alcaligenes genus microorganism, Alteromonas genus microorganism, Amoebobacter genus microorganism, Aphanocapa sp., Aphanothece sp. Aquaspirillum autotrophicum, Azorhizobium caulinodans, Azospirillum sp ., Azospirillum spp, Azotobacter spp, Bacillus spp, Beggiatoa spp, Beijerinckia spp, Beneckea spp, Bordetella pertussis, Bradyrhizobium japonicum, Caryophamon latum, Caulobacter spp, Chlorogloea spp ., Chromatium microorganism of the genus, Chromobacterium spp, Clostridium spp, Comamonas spp, Corynebacterium spp, Cyanobacteria in microorganisms, Derxia spp, Desulfonema spp, Desulfosacina variabilis, Desulfovibrio sapovorans, Ectothiorhodospira spp, Ferrobacillus ferroxidans, Flavobacterium sp, Haemophilus influenzae, Halobacterium spp, Haloferax mediterranei, Hydroclathratus clathratus, Hydrogenomonas facilis, microorganisms in Hydrogenophaga, microorganisms in Hyphomicrobium, Ilyobacter delafieldii, Labrys monachus, Lamprocystis reseopersicina, Lampropedia hyalina, Legionella sp., L eptothrix discophorus, Methylobacterium spp, Methylosinus spp, Micrococcus spp, Mycobacterium spp, Nitrobacter spp, Nocardia spp, Paracoccus dentrificans, Oscillatoria limosa, Penicillium cyclopium, Photobacterium spp, Physarum ploycephalum, Pseudomonas spp, Ralstonia spp , Rhizobium spp, Rhodobacillus spp, Rhodobacter spp, Rhodococcus genus microorganisms, Rhodocyclus spp, Rhodomicrobium vannielii, Rhodopseudomonas spp, Rhodospirillum spp, Sphingomonas paucimobilis, Spirillum spp, Spirulina spp, Staphylococcus spp, Stella in microorganism, Streptomyces spp, Syntrophomonas wolfei, Thermophilic cyanobacteria, Thermus thermophilus, Thiobacillus A2, Thiobacillus spp, Thiocapsa spp, Thiocystis violacea, Vibrio parahaemolyticus, Xanthobacter autotrophicus, Xanthomonas maltophilia, Zoogloea And a method for producing poly-3-hydroxybutyrate 3-hydroxybutyrate methyl ester, characterized in that the enzyme is selected from the group consisting of a microorganism transformed with a gene encoding having a producing ability. 제14항에 있어서, 폴리-3-하이드록시부티레이트{poly(3-hydroxybutyrate)} 생성능을 가지는 미생물은 Alacligenes latus인 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법. 15. The method of claim 14, wherein the microorganism having a poly-3-hydroxybutyrate production ability is Alacligenes latus . 제13항에 있어서, (a) 단계의 배양은 질소원이 제한된 배지에서 수행하는 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The method for preparing 3-hydroxybutyrate methyl ester according to claim 13, wherein the culturing of step (a) is performed in a medium with a limited nitrogen source. 제13항에 있어서, (c) 단계의 반응은 80~120℃에서 1~24시간 수행하는 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The method of claim 13, wherein the reaction of step (c) is performed at 80 to 120 ° C. for 1 to 24 hours. 제15항에 있어서, 유기용매는 클로로포름인 것을 특징으로 하는 3-하이드록시부티레이트 메틸에스테르의 제조방법.The process for producing 3-hydroxybutyrate methyl ester according to claim 15, wherein the organic solvent is chloroform.
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US9102972B2 (en) 2012-03-05 2015-08-11 Korea Research Institute Of Bioscience And Biotechnology Microorganism Rhizobium sp. KB10 having properties of promoting growth of Botryococcus braunii and increasing fatty acid content
CN104830709A (en) * 2015-02-16 2015-08-12 南京大学 High-temperature-resistant aerobic denitrification bacterial strain and application of same
CN112940969A (en) * 2021-02-07 2021-06-11 兴安盟莱绅生物农业有限公司 Novel microbial agent and soybean planting method

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* Cited by examiner, † Cited by third party
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US9102972B2 (en) 2012-03-05 2015-08-11 Korea Research Institute Of Bioscience And Biotechnology Microorganism Rhizobium sp. KB10 having properties of promoting growth of Botryococcus braunii and increasing fatty acid content
CN104830709A (en) * 2015-02-16 2015-08-12 南京大学 High-temperature-resistant aerobic denitrification bacterial strain and application of same
CN112940969A (en) * 2021-02-07 2021-06-11 兴安盟莱绅生物农业有限公司 Novel microbial agent and soybean planting method

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