KR101311007B1 - Peracetic acid production method and acetyl xylan esterase for peracetic acid production - Google Patents

Peracetic acid production method and acetyl xylan esterase for peracetic acid production Download PDF

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KR101311007B1
KR101311007B1 KR1020110118777A KR20110118777A KR101311007B1 KR 101311007 B1 KR101311007 B1 KR 101311007B1 KR 1020110118777 A KR1020110118777 A KR 1020110118777A KR 20110118777 A KR20110118777 A KR 20110118777A KR 101311007 B1 KR101311007 B1 KR 101311007B1
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peracetic acid
acetyl xylan
xylan esterase
acid production
present
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KR20130053573A (en
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박승문
김대혁
권태호
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(주)엔비엠
전북대학교산학협력단
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/54—Acetic acid
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • C12N9/18—Carboxylic ester hydrolases (3.1.1)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y301/00—Hydrolases acting on ester bonds (3.1)
    • C12Y301/01—Carboxylic ester hydrolases (3.1.1)
    • C12Y301/01072—Acetylxylan esterase (3.1.1.72)

Abstract

본 발명은 과아세트산 생산방법에 관한 발명이다. 또한 본 발명은 과아세트산 생산용 아세틸 자일란 에스터라제에 관한 발명이다. 이러한 본 발명은 아세틸 자일란 에스터라제에 에틸 아세테이트 및 과산화수소를 처리하게 되면 아세틸 자일란 에스터라제의 기존 효과인 탈아세틱화 반응을 촉매하는 효소로서의 기능에 더하여, 과아세트산을 생산할 수 있게 된다. 그리하여 본 발명에 따라 아세틸 자일란 에스터라제에 의해 과아세트산을 생산하게 되면 과아세트산의 생산 경로를 보다 다양화 할 수 있게 되어 과아세트산의 생산 비용을 낮출 수 있을 것으로 기대된다. 또한 본 발명은 아세틸 자일란 에스터라제에 대하여 과아세트산을 생산하는 새로운 용도를 밝힌 발명이다. The present invention relates to a method for producing peracetic acid. The invention also relates to an acetyl xylan esterase for peracetic acid production. In the present invention, the treatment of acetyl xylan esterase with ethyl acetate and hydrogen peroxide can produce peracetic acid in addition to its function as an enzyme catalyzing the deacetylation reaction, which is an existing effect of acetyl xylan esterase. Therefore, when the peracetic acid is produced by the acetyl xylan esterase according to the present invention, it is expected that the production route of the peracetic acid can be more diversified, thereby lowering the production cost of the peracetic acid. The present invention also discloses a novel use of acetyl xylan esterase to produce peracetic acid.

Description

과아세트산 생산 방법 및 과아세트산 생산용 아세틸 자일란 에스터라제{Peracetic acid production method and acetyl xylan esterase for peracetic acid production} Peracetic acid production method and acetyl xylan esterase for peracetic acid production}

본 발명은 과아세트산 생산 방법 및 과아세트산 생산용 아세틸 자일란 에스터라제에 관한 것이다.
The present invention relates to a method for producing peracetic acid and to acetyl xylan esterases for producing peracetic acid.

아세틸 자일란 에스터라제(Acetyl xylan esterase)는 자일란이나 자일로 올리고 사카라이드의 탈아세틸화 반응을 촉매하는 효소로 알려져 있다. 특히 아세틸 자일란 에스터라제는 자일란 아세틸기의 에스터 결합을 가수분해 시키는 효소로서, 헤미셀룰로스를 구성하는 자일란(Xylan) 구조에 위치하는 아세틸 그룹 (acetyl group)을 제거하는 효소로 알려져 있다.Acetyl xylan esterase is known as an enzyme that catalyzes the deacetylation reaction of xylan or xylo oligosaccharides. In particular, acetyl xylan esterase is an enzyme that hydrolyzes the ester bond of the xylan acetyl group, and is known as an enzyme that removes an acetyl group located in the xylan structure constituting hemicellulose.

또한 과아세트산(Peracetic acid, PAA)은 강력한 산화기능이 있는 물질로서 멸균 소독 등에 사용되는 산화제로서, 특히 바이오매스에서 리그닌을 산화시킨다. 이렇게 산화된 리그닌은 분해되므로, 결과적으로 과아세트산은 바이오매스의 사전처리를 가능하게 하는 화합물인 것이다. 이렇게 과아세트산으로 사전 처리된 바이오매스는 그 효율이 보다 향상되게 되고, 보다 효율적으로 에탄올 등을 생산해 낼 수 있는 것이다. 또한 이러한 과아세트산은 제지 산업에서 표백제로도 사용될 수 있다. 그러므로 과아세트산은 바이오매스 및 제지 산업에 유용하게 쓰이는 화합물이다.
In addition, peracetic acid (PAA) is a powerful oxidizing material used for sterilization and disinfection, especially oxidizing lignin in biomass. This oxidized lignin is decomposed, and consequently, peracetic acid is a compound that allows pretreatment of biomass. The biomass pretreated with peracetic acid is thus more efficient and can produce ethanol and the like more efficiently. Such peracetic acid can also be used as bleach in the paper industry. Peracetic acid is therefore a compound useful for the biomass and paper industry.

상기와 같은 종래기술에 더하여 본 발명에 따른 과아세트산 생산방법은 바이오매스 및 제지 산업에 쓰이는 과아세트산을 아세틸 자일란 에스터라제를 이용하여 생산하고자 하는 것이다. 또한, 본 발명에 따른 과아세트산 생산용 아세틸 자일란 에스터라제는 탈아세틸화 반응을 촉매하는 효소로 알려져 있던 아세틸 자일란 에스터라제에 대해 과아세트산을 생산하는 새로운 용도가 있음을 밝혀 그 활용도를 보다 다양화하고자 하는 것이다.
In addition to the prior art as described above, the method for producing peracetic acid according to the present invention is to produce peracetic acid used in the biomass and paper industry using acetyl xylan esterase. In addition, the acetyl xylan esterase for producing peracetic acid according to the present invention has been found to be a new use for producing peracetic acid for acetyl xylan esterase known as an enzyme catalyzing the deacetylation reaction, the utilization of the It is to be made.

위와 같은 과제를 해결하기 위한 본 발명의 한 특징에 따른 과아세트산 생산 방법은 아세틸 자일란 에스터라제를 이용하여 과아세트산을 생산하는 것에 관한 것이다. 또한 상기 아세틸 자일란 에스터라제에는 에틸 아세테이트 및 과산화수소가 처리되어 과아세트산을 생산하는 과아세트산 생산 방법에 관한 것이다.Peracetic acid production method according to a feature of the present invention for solving the above problems relates to the production of peracetic acid using acetyl xylan esterase. The acetyl xylan esterase also relates to a method for producing peracetic acid in which ethyl acetate and hydrogen peroxide are treated to produce peracetic acid.

본 발명의 또 다른 특징에 따른 아세틸 자일란 에스터라제는 과아세트산 생산용 아세틸 자일란 에스터라아제에 관한 것이다. 또한 상기 과아세트산 생산은 에틸 아세테이트와 과산화수소가 처리되어 생산되는 것을 특징으로 하는 과아세트산 생산용 아세틸 자일란 에스터라제에 관한 것이다.
Acetyl xylan esterase according to another feature of the present invention relates to an acetyl xylan esterase for peracetic acid production. In addition, the peracetic acid production relates to acetyl xylan esterase for peracetic acid production, characterized in that the ethyl acetate and hydrogen peroxide are processed.

본 발명은 과아세트산 생산방법에 관한 발명으로서, 바이오매스 산업이나 제지산업에 널리 쓰이는 과아세트산의 생산 경로를 보다 다양화 할 수 있게 되어 그 공급을 보다 원활하게 할 수 있다. 그리하여 과아세트산의 공급 가격을 낮출 수 있을 것으로 기대된다. 또한 아세틸 자일란 에스터라제의 효과 중 탈아세틸화 반응을 촉매하는 효소로서의 효과에 더하여, 과아세트산을 생산하는 효과를 추가로 발견한 것으로서 아세틸 자일란 에스터라제의 활용도를 다양화한 발명이다.
The present invention relates to a method for producing peracetic acid, and the production route of peracetic acid, which is widely used in the biomass industry and the paper industry, can be more diversified, and the supply thereof can be made smoother. Thus, it is expected that the supply price of peracetic acid can be lowered. In addition to the effects of acetyl xylan esterase as an enzyme catalyzing the deacetylation reaction, the inventors further discovered the effect of producing peracetic acid, and is an invention in which the utilization of acetyl xylan esterase is diversified.

도 1은 본 발명의 실시예에 따라 분리된 아세틸 자일란 에스터라제의 SDS-PAGE 분석 결과를 나타낸 사진이다.
도 2는 본 발명의 실시예에 따른 MTS의 산화물을 확인하기 위한 HPLC로 측정한 결과를 나타낸 그래프이다.
도 3은 반응시간에 따른 MTSO와 MTSON의 증감 정도를 관찰한 결과를 나타낸 그래프이다. 1 is a photograph showing the results of SDS-PAGE analysis of acetyl xylan esterase isolated according to an embodiment of the present invention.
Figure 2 is a graph showing the results measured by HPLC for confirming the oxide of the MTS according to an embodiment of the present invention.
3 is a graph showing the results of observing the degree of increase and decrease of MTSO according to the reaction time.

이에 본 발명자들은 과아세트산의 생산 경로를 다양화 하고, 아세틸 자일란 에스터라제의 새로운 용도를 발견하기 위해 예의 연구 노력한 결과, 과아세트산 생산방법 및 과아세트산 생산용 아세틸 자일란 에스터라제에 관한 본 발명을 완성하였다.
Accordingly, the present inventors have made extensive efforts to diversify the production route of peracetic acid and to discover new uses of acetyl xylan esterase. As a result, the present invention relates to a method for producing peracetic acid and acetyl xylan esterase for peracetic acid production. Completed.

구체적으로는 본 발명에 따른 과아세트산 생산방법은 아세틸 자일란 에스터라제를 이용하여 과아세트산을 생산하는 방법에 관한 것이다. Specifically, the method for producing peracetic acid according to the present invention relates to a method for producing peracetic acid using acetyl xylan esterase.

또한 본 발명에 따른 과아세트산 생산방법은 에틸 아세테이트(ethyl acetate) 및 과산화수소(hydrogen peroxide)를 아세틸 자일란 에스터라제에 처리하게 되면 에틸 아세테이트가 과아세틱산으로 전환되어 과아세틱산을 생산하게 되는 것이다. In addition, the method of producing peracetic acid according to the present invention is that when ethyl acetate and hydrogen peroxide are treated with acetyl xylan esterase, ethyl acetate is converted to peracetic acid to produce peracetic acid.

상기 아세틸 자일란 에스터라제는 재조합 아세틸 자일란 에스터라제 유전자에 의해 발현된 것을 포함한다. 또한 상기 아세틸 자일란 에스터라제는 이를 생산하는 균주에 의해 발현된 것이 포함 된다.
The acetyl xylan esterases include those expressed by recombinant acetyl xylan esterase genes. In addition, the acetyl xylan esterase includes that expressed by the strain producing it.

또한 본 발명에 따른 과아세트산 생산용 아세틸 자일란 에스터라제는 아세틸 자일란 에스터라제에 에틸 아세테이트(ethyl acetate) 및 과산화수소(hydrogen peroxide)를 처리하게 되면 과아세트산이 생산되는 것으로서, 아세틸 자일란 에스터라제의 새로운 용도에 관한 발명이다. In addition, the acetyl xylan esterase for producing peracetic acid according to the present invention is treated with acetyl xylan esterase and ethyl acetate (ethyl acetate) and hydrogen peroxide (hydrogen peroxide) to produce peracetic acid, the acetyl xylan esterase The invention relates to a new use.

상기 아세틸 자일란 에스터라제는 재조합 아세틸 자일란 에스터라제 유전자에 의해 발현된 것을 포함한다. 또한 상기 아세틸 자일란 에스터라제는 이를 생산하는 균주에 의해 발현된 것이 포함 된다.
The acetyl xylan esterases include those expressed by recombinant acetyl xylan esterase genes. In addition, the acetyl xylan esterase includes that expressed by the strain producing it.

이하 본 발명을 바람직한 실시예를 참고로 하여 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있도록 상세히 설명한다. 그러나 본 발명은 여러 가지 상이한 형태로 구현될 수 있으며 여기에서 설명하는 실시예에 한정되는 것은 아니다.
Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. The present invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

실시예Example

<< 실시예Example 1: 아세틸  1: acetyl 자일란Xylan 에스터라제의Esterase 분리> Separation>

Aspergillus ficumm의 아세틸 자일란 에스터라제를 코딩하는 유전자를 Pichia pastoris 의 pPICZc 부위에 삽입하여 재조합 아세틸 자일란 에스터라제 유전자를 제조하였다. 그리고 이러한 재조합 아세틸 자일란 에스터라제 유전자를 클로닝하였으며, 전기천공법 (electrophoretion)을 통하여 이러한 클로닝 된 유전자를 포함하는 Pichia pastoris GS115(his4)의 형질전환체(transformant)를 얻었다. 이렇게 얻은 형질전환체는 PDA 배지에 접종하였으며, 이를 배양하여 보관하였다. A gene encoding the acetyl xylan esterase of Aspergillus ficumm was inserted into the pPICZc region of Pichia pastoris to prepare a recombinant acetyl xylan esterase gene. Then, the recombinant acetyl xylan esterase gene was cloned, and a transformant of Pichia pastoris GS115 (his4) containing the cloned gene was obtained through electrophoretion. The transformants thus obtained were inoculated in PDA medium, and then cultured and stored.

그 후 이러한 형질전환체를 탄소원으로서 글리세롤이 첨가된 100 mL BMGY 배지에 소량 접종하고, 30℃에서 진탕 배양 하였다. 배양 중 현탁액의 흡광도가 600 nm에서 6.0에 이르면 원심분리하여 펠렛 만을 회수하였고, 펠렛에 잔존하고 있는 글리세롤은 멸균 증류수로 2회 세척하여 제거하였다. 세척된 펠렛 만을 0.5% 메탄올이 첨가된 100 mL BMMY 배지에 현탁하고 진탕배양하여 아세틸 자일란 에스터라제가 발현되도록 유도하였다. 이를3일 정도 배양 후, 배양현탁액을 원심분리하여 배양 상등액 만을 회수하였다. Ni+2 His tag column (Histrap, GE Healthcare)을 이용하여 배양 상등액에서 재조합 아세틸 자일란 에스터라제 유전자에 의해 발현된 아세틸 자일란 에스터라제 만을 분리하였고, 분리 중에 첨가된 염(salt)과 이미다졸(imidazole)은 투석을 통해 제거하였으며, 아세틸 자일란 에스터라제의 농도는 브레드포드(Bradford) 방법으로 측정하였다. 이후 최종적으로 분리된 아세틸 자일란 에스터라제는 SDS-PAGE(Sodium dodecyl sulfate polyacrylamide gel electrophoresis)로 확인하였다. 도 1의 M은 이러한 SDS-PAGE의 결과를 나타낸 것이며, 레인 1은 P. pastoris에서 분리된 아세틸 자일란 에스터라제의 분자량을 확인한 결과를 나타낸다.
Thereafter, these transformants were inoculated in small amounts in 100 mL BMGY medium to which glycerol was added as a carbon source, followed by shaking culture at 30 캜. When the absorbance of the suspension reached 600 to 6.0 at 600 nm, the pellets were recovered by centrifugation. Glycerol remaining in the pellets was removed by washing twice with sterile distilled water. Only washed pellets were suspended in 100 mL BMMY medium with 0.5% methanol and shaken to induce acetyl xylan esterase expression. After culturing for about 3 days, the culture suspension was centrifuged to recover only the culture supernatant. Only the acetyl xylan esterase expressed by the recombinant acetyl xylan esterase gene was isolated from the culture supernatant using a Ni + 2 His tag column (Histrap, GE Healthcare), and the salt and imidazole added during the separation ( imidazole) was removed by dialysis, and the concentration of acetyl xylan esterase was measured by the Bradford method. After the final acetyl xylan esterase was confirmed by SDS-PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis). Figure 1 M shows the results of this SDS-PAGE, lane 1 shows the result of confirming the molecular weight of acetyl xylan esterase isolated from P. pastoris.

<< 실시예Example 2: 분리한 아세틸  2: separated acetyl 자일란Xylan 에스터라제의Esterase 과아세트산Peracetic acid 생산 여부 조사> Investigate Production

실시예 1로 얻은 아세틸 자일란 에스터라제의 가수분해(perhydrolysis) 활성은 술폭시화물(sulfoxide) 생성 여부를 HPLC를 통해 정량하여 측정하였다. 측정방법은 100 mM 포타시움 포스페이트(potassium phosphate) 완충용액(pH 7.0)에 500mM의 에틸 아세테이트(ethyl acetate), 1M의 과산화수소(hydrogen peroxide) 및 아세틸 자일란 에스터라제를 첨가하였다. 이때 반응액의 부피는 1 mL로 37℃에서 30분간 반응하였다. 반응 후, 반응액에서 0.1 mL 만을 취하여 메틸 p-tolyl 설파이드(methyl p-tolyl sulfide, MTS, 20 mM) 0.1 mL 그리고 60% 아세톤니트릴(acetonitrile) 0.8 mL과 혼합하여 37℃에서 30분간 반응하였다. 10 mg의 MnO2를 반응액에 첨가하여 반응을 정지시키고 잔존하고 있는 MnO2는 원심분리하여 제거하였다. 잔존하고 있는 MTS 양과 아세틸 자일란에 의한 가수분해(perhydrolysis) 반응으로 생성된 methyl p-tolyl sulfoxide (MTSO)와 methyl p-tolyl sulfone (MTSON)과 같은 MTS의 산화물의 양은 HPLC를 이용하여 정량 그리고 정성 분석하였다. 또한 HPLC/MS로 MTS의 가수분해(perhydrolysis) 결과로 MTSO와 MTSON의 생성 유무를 검증하였다. 그리하여 아세틸 자일란 에스터라제가 과산화수소 존재하에 에틸 아세테이트(ethyl acetate)를 과아세트산(PAA)으로 전환시키는 가수분해(perhydrolysis) 활성을 지닌 것을 알 수 있었다.
The perhydrolysis activity of the acetyl xylan esterase obtained in Example 1 was determined by quantitative determination of sulfoxide production through HPLC. In the measurement method, 500 mM ethyl acetate, 1 M hydrogen peroxide and acetyl xylan esterase were added to 100 mM potassium phosphate buffer (pH 7.0). At this time, the reaction solution was reacted at 37 ° C. for 30 minutes at 1 mL. After the reaction, only 0.1 mL of the reaction solution was taken and mixed with 0.1 mL of methyl p-tolyl sulfide (MTS, 20 mM) and 0.8 mL of 60% acetonitrile and reacted at 37 ° C. for 30 minutes. 10 mg of MnO 2 was added to the reaction solution to stop the reaction, and the remaining MnO 2 was removed by centrifugation. Quantification and qualitative analysis of the amount of MTS remaining and the amount of MTS oxides such as methyl p-tolyl sulfoxide (MTSO) and methyl p-tolyl sulfone (MTSON) produced by perhydrolysis by acetyl xylan It was. In addition, the presence or absence of MTSO and MTSON was verified by perhydrolysis of MTS by HPLC / MS. Thus, it was found that acetyl xylan esterase has a perhydrolysis activity to convert ethyl acetate to peracetic acid (PAA) in the presence of hydrogen peroxide.

<< 실시예Example 3: 아세틸  3: acetyl 자일란Xylan 에스터라제에Esterase 의해 전환되는  Being switched by 과아세트산의Peracetic acid 양> Quantity>

가수분해(Perhydrolysis)로 생성되는 과아세트산(PAA)의 양은 MTS 산화로 간접적으로 측정할 수 있었다. MTS의 산화물은 HPLC로 측정하였다. HPLC로 분석한 결과 네 개의 피크를 확인 할 수 있었는데, 이 중 피크 A와 피크 B는 각각 과아세트산에 의한 MTS 산화로 인해 형성된 MTSO 그리고 MTSON임을 알 수 있었다. 이의 결과는 도 2에 나타냈다. 또한 반응시간에 따른 MTSO와 MTSON의 증감 정도를 관찰하였으며 이의 결과를 도 3에 나타냈다. 구체적으로는 과아세트산(PAA)에 의한 MTS 산화의 첫 번째 산물로 MTSO가 생성되었으며, 반응시간이 지속됨에 따라 MTSO가 재차 산화되어 MTSO의 양은 줄어든 반면 MTSON이 생성됨을 알 수 있었다. 120분 동안 지속된 과아세트산(PAA)에 의한 MTS의 산화로 MTSO와 MTSON는 0.72 mM과 0.31 mM이 각각 생성됨을 알 수 있었다. 이러한 결과는 10ug의 아세틸 자일란 에스터라제에 의해 1.34mM의 과아세트산(PAA)이 생성됨을 의미한다.
The amount of peracetic acid (PAA) produced by hydrolysis could be measured indirectly by MTS oxidation. The oxide of MTS was measured by HPLC. As a result of HPLC analysis, four peaks were identified, and peak A and peak B were MTSO and MTSON formed by MTS oxidation by peracetic acid, respectively. The results are shown in FIG. In addition, the degree of increase and decrease of MTSO and MTSON was observed according to the reaction time, and the results are shown in FIG. 3. Specifically, MTSO was produced as the first product of MTS oxidation by peracetic acid (PAA). As the reaction time was continued, MTSO was oxidized again to reduce MTSO content while producing MTSON. MTSO and MTSON produced 0.72 mM and 0.31 mM, respectively, by oxidation of MTS by 120% peracetic acid (PAA). This result means that 1.34 mM peracetic acid (PAA) is produced by 10 ug of acetyl xylan esterase.

상기에서는 본 발명의 바람직한 실시예에 대하여 설명하였지만, 본 발명은 이에 한정되는 것은 아니고, 본 발명의 기술 사상 범위 내에서 여러 가지로 변형하여 실시하는 것이 가능하고, 이 또한 첨부된 특허 청구 범위에 속하는 것은 당연하다.
While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims. It is natural.

Claims (4)

과산화수소하에서, 에틸아세테이트를 아세틸 자일란 에스터라제로 가수분해하여 과아세트산이 제조되며,
상기 아세틸 자일란 에스터라제는 진균 아스펄질러스 피컴(Aspergillus ficuum) 유래의 효소인 것을 특징으로 하는 과아세트산 생산 방법.
Under hydrogen peroxide, ethyl acetate is hydrolyzed with acetyl xylan esterase to prepare peracetic acid,
The acetyl xylan esterase is a peracetic acid production method, characterized in that the enzyme derived from the fungus Aspergillus ficuum.
제 1항에 있어서,
상기 아세틸 자일란 에스터라제는 진균 아스펄질러스 피컴(Aspergillus ficuum) 유래의 유전자를 효모 피키아 파스토리스(Pichia pastoris) GS115(his4)에 형질전환시켜 형질전환체(transformant)로부터 생산되는 것을 특징으로 하는 과아세트산 생산방법.
The method of claim 1,
The acetyl xylan esterase is produced from a transformant by transforming a gene derived from the fungus Aspergillus ficuum into yeast Pichia pastoris GS115 (his4). Peracetic acid production method.
제 2항에 있어서,
상기 아세틸 자일란 에스터라제는 분자량이 29.5 ~ 34 kDa인 것을 특징으로 하는 과아세트산 생산방법.
The method of claim 2,
The acetyl xylan esterase is a peracetic acid production method, characterized in that the molecular weight of 29.5 ~ 34 kDa.
제 1항에 있어서,
10ug의 아세틸 자일란 에스터라제에 의해 1.34mM의 과아세트산(PAA)을 생산하는 것을 특징으로 하는 과아세트산 생산방법.The method of claim 1,
A method for producing peracetic acid, characterized by producing 1.34 mM peracetic acid (PAA) by 10 ug of acetyl xylan esterase.
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