KR100318755B1 - Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization - Google Patents
Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization Download PDFInfo
- Publication number
- KR100318755B1 KR100318755B1 KR1019990061537A KR19990061537A KR100318755B1 KR 100318755 B1 KR100318755 B1 KR 100318755B1 KR 1019990061537 A KR1019990061537 A KR 1019990061537A KR 19990061537 A KR19990061537 A KR 19990061537A KR 100318755 B1 KR100318755 B1 KR 100318755B1
- Authority
- KR
- South Korea
- Prior art keywords
- ethanol
- wood
- producing
- temperature sterilization
- glucose
- Prior art date
Links
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 title claims abstract description 144
- 230000001954 sterilising effect Effects 0.000 title claims abstract description 47
- 239000002023 wood Substances 0.000 title claims abstract description 47
- 238000004659 sterilization and disinfection Methods 0.000 title claims abstract description 45
- 238000000034 method Methods 0.000 title claims abstract description 40
- 239000000413 hydrolysate Substances 0.000 title 1
- 238000000855 fermentation Methods 0.000 claims abstract description 47
- 230000004151 fermentation Effects 0.000 claims abstract description 47
- 238000004519 manufacturing process Methods 0.000 claims abstract description 25
- 239000002028 Biomass Substances 0.000 claims abstract description 23
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 239000010815 organic waste Substances 0.000 claims abstract description 9
- 238000009928 pasteurization Methods 0.000 claims abstract 4
- 239000000243 solution Substances 0.000 claims description 34
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 29
- 239000008103 glucose Substances 0.000 claims description 29
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 22
- 244000005700 microbiome Species 0.000 claims description 13
- 239000003440 toxic substance Substances 0.000 claims description 11
- 231100000614 poison Toxicity 0.000 claims description 10
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 8
- 230000000694 effects Effects 0.000 claims description 8
- 238000011282 treatment Methods 0.000 claims description 8
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 7
- 210000004027 cell Anatomy 0.000 claims description 7
- 238000001914 filtration Methods 0.000 claims description 7
- 239000002699 waste material Substances 0.000 claims description 7
- 241000235070 Saccharomyces Species 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 5
- 239000000919 ceramic Substances 0.000 claims description 3
- 239000011148 porous material Substances 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 210000005253 yeast cell Anatomy 0.000 claims description 2
- 238000007429 general method Methods 0.000 abstract description 2
- 238000010170 biological method Methods 0.000 abstract 1
- 238000000746 purification Methods 0.000 abstract 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 23
- 239000000126 substance Substances 0.000 description 9
- 239000000463 material Substances 0.000 description 5
- 230000002255 enzymatic effect Effects 0.000 description 3
- 108010047754 beta-Glucosidase Proteins 0.000 description 2
- 102000006995 beta-Glucosidase Human genes 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 108010023063 Bacto-peptone Proteins 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 238000012365 batch cultivation Methods 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229940106157 cellulase Drugs 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
-
- 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
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
-
- 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
- C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
- C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
- C12P7/06—Ethanol, i.e. non-beverage
- C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
- C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
본 발명은 저온멸균법을 이용한 목질계 당화액으로부터 고농도 에탄올 제조 방법에 관한 것으로, 그 목적은 목질계 바이오매스로부터 에탄올을 생산하는데 있어서 기존 공정에서 사용하는 고온 멸균대신에, 저온 멸균법을 이용하여 일반적인 방법에 의해 생산 가능한 에탄올 농도 이상을 얻을 뿐만 아니라 생산성이 높은 회분식 및 연속식 목질계 에탄올 생산공정을 제공하는데 있다.The present invention relates to a method for producing a high concentration of ethanol from wood-based saccharification liquid using a pasteurization method, the purpose of which is a general method using low-temperature sterilization instead of the high-temperature sterilization used in the existing process in the production of ethanol from wood-based biomass In addition to obtaining more than the ethanol concentration that can be produced by the present invention to provide a high productivity batch and continuous wood-based ethanol production process.
본 발명의 제공에 의하여 에탄올을 고농도, 고생산성으로 생산할 수 있어 에탄올 생산 전체 공정 중 발효 및 정제에 소요되는 비용을 낮출 수 있는데, 본 발명의 회분식 및 연속식 발효에 의하면, 에탄올 농도와 생산성이 각각 67 g/L, 1.0 g/L/h 와, 76.9 g/L, 16.9 g/L/h를 달성할 수 있었다.Providing the present invention can produce ethanol at high concentration and high productivity, thereby reducing the cost of fermentation and purification during the entire ethanol production process. According to the batch and continuous fermentation of the present invention, ethanol concentration and productivity are respectively 67 g / L, 1.0 g / L / h, and 76.9 g / L, 16.9 g / L / h could be achieved.
본 방법은 전세계적으로 풍부하게 있고 환경 측면에서도 중요한 자원인 목재, 도시유기성 폐기물 등 목질계 바이오매스를 이용하여 환경 친화적인 생물학적 방법을 통하여 높은 농도 및 생산성으로 에탄올을 생산할 수 있기 때문에 환경 및 경제적 측면에서 장점을 갖는 공정이다.This method is environmentally and economically because it can produce ethanol at high concentration and productivity through environmentally friendly biological methods using wood-based biomass such as wood and municipal organic waste, which are abundant in the world and are important resources in the environment. It is a process with advantages.
Description
본 발명은 저온멸균법을 이용한 목질계 당화액으로부터 고농도 에탄올 제조 방법에 관한 것으로, 상세하게 설명하자면 나무, 폐목재, 도시유기성 폐기물의 당화액에 존재하는 포도당으로부터 효모인사카로마이세스 세레비제(Saccharomyces cerevisiae)를 이용하여 에탄올을 생산하는데 있어서, 기존의 멸균법과는 다른 저온 멸균을 통해 목질계 바이오매스(cellulosic biomass)로부터 생기는 미생물에 유해한 여러 독성물질의 생성을 억제하도록 하고 상기 효모를 이용하여 목질계 바이오매스로부터 고농도로 에탄올을 생산하는 방법에 관한 것이다.The present invention relates to a high-concentration ethanol production process from lignocellulosic Hydrolyzate with low temperature myeolgyunbeop, described in detail gritty wood, waste wood, in yeast Saccharomyces from the glucose present in the saccharification mixture urban organic waste, my process serenity non-zero (Saccharomyces cerevisiae) in the production of ethanol, by the low temperature sterilization different from the conventional sterilization method to suppress the production of various toxic substances harmful to microorganisms generated from cellulosic biomass and the yeast system A method for producing ethanol at high concentration from biomass.
목질계 바이오매스는 환경친화성과 값싸고 풍부하게 존재한다는 이유로 인해 이들 물질을 이용하여 에탄올 등 유용한 물질을 생산하고자 하는 연구가 최근에 관심을 끌고 있다(참조: Lynd et al., Science, 251: 1318-1323 (1991)).Woody biomass has recently attracted interest in producing useful substances such as ethanol using these materials due to their environmental friendliness and inexpensive and abundant existence (Lynd et al., Science, 251: 1318). -1323 (1991)).
미생물을 이용하여 목질계 바이오매스로부터 유용한 물질을 생산하기 위해서는 포도당 등 미생물이 이용할 수 있는 형태의 당으로 목질계 바이오매스를 전처리하는 것이 필요하다.In order to produce useful substances from woody biomass using microorganisms, it is necessary to pretreat woody biomass with sugars in the form of microorganisms such as glucose.
그러나 목질계 바이오 매스를 전처리하는 과정에서 많은 종류의 미생물의 성장에 나쁜 독성물질들이 생성된다.However, pretreatment of wood-based biomass produces toxic substances that are bad for the growth of many microorganisms.
일반적으로 미생물을 이용하여 에탄올 등 유용물질을 생산하는 경우 정제 등에 드는 비용을 고려할 때 미생물로부터 생산된 에탄올 등 유용물질의 농도가 높을수록 경제성이 있다.In general, in the case of producing useful substances such as ethanol using microorganisms, the higher the concentration of useful substances such as ethanol produced from microorganisms, the more economical the economical cost is.
그러나 목질계 바이오매스를 이용하는 경우, 이러한 독성물질들로 인해 에탄올 등의 유용물질을 고농도로 생산할 수 없었다.However, when using wood-based biomass, these toxic substances could not produce high concentrations of useful substances such as ethanol.
그리하여 이들 독성물질들을 활성탄, 석회(lime) 등을 이용하여 화학적으로 처리하여 독성을 줄이는 연구가 있어 왔다(Parajo et al., Bioresource Technol., 57: 179-185 (1996), Rivard et al., Appl. Biochem. Biotechnol., 57-58: 183-191 (1996)).Thus, there have been studies to reduce the toxicity by chemically treating these toxic substances with activated carbon, lime, etc. (Parajo et al., Bioresource Technol., 57: 179-185 (1996), Rivard et al., Appl. Biochem.Biotechnol., 57-58: 183-191 (1996)).
그러나 이들 화학적 처리는 첨가 물질의 분리 등 또 다른 문제점을 야기하고 있으며 또한 그 처리 능력도 물질, 방법에 따라 차이가 있어 목질계 바이오 매스로부터 경제적으로 에탄올을 생산하기 위해서는 이들 문제점을 해결할 수 있는 또 다른 방법의 개발이 필요하였다.However, these chemical treatments cause other problems, such as separation of additive materials, and the processing capacity varies depending on the materials and methods, so that these problems can be solved in order to economically produce ethanol from woody biomass. Development of the method was needed.
또한 에탄올 등 범용 화학제품을 목질계 바이오매스로부터 경제적으로 생산하기 위해서는 생산된 에탄올 농도뿐만 아니라 발효기의 생산성도 매우 중요하여 효율적인 발효공정의 개발도 필요하다.In addition, in order to economically produce general-purpose chemicals such as ethanol from woody biomass, not only the produced ethanol concentration but also the productivity of the fermenter is very important, and therefore, it is necessary to develop an efficient fermentation process.
본 발명의 목적은 상기와 같은 문제점을 해결하고자 목질계 바이오매스로부터 에탄올을 생산하는데 있어서 기존 공정에서 사용하는 고온 멸균대신에, 저온 멸균법을 이용하여 일반적인 방법에 의해 생산 가능한 에탄올 농도 이상을 얻을 뿐만 아니라 생산성도 높게 에탄올을 생산할 수 있는 회분식 및 연속식 에탄올 발효 공정을 제공하는데 있다.It is an object of the present invention to obtain ethanol concentration that can be produced by a general method using low temperature sterilization instead of high temperature sterilization in the conventional process in producing ethanol from woody biomass to solve the above problems. It is to provide a batch and continuous ethanol fermentation process capable of producing ethanol with high productivity.
도 1은 고온 멸균시(121℃) 목질계 당화액으로부터 에탄올 발효능력을 나타낸 그래프이다.1 is a graph showing the ethanol fermentation capacity from the wood-based saccharification solution at high temperature sterilization (121 ℃).
도 2는 활성탄 처리 후 고온 멸균시(121℃) 목질계 당화액으로부터 에탄올 발효능력을 나타낸 그래프이다.Figure 2 is a graph showing the ethanol fermentation capacity from the wood-based saccharification solution at high temperature sterilization (121 ℃) after activated carbon treatment.
도 3은 저온 멸균시(60℃) 목질계 당화액으로부터 에탄올 발효능력(a: 포도당 농도, b: 에탄올 농도)을 나타낸 그래프이다.Figure 3 is a graph showing the ethanol fermentation capacity (a: glucose concentration, b: ethanol concentration) from the wood-based saccharification solution at low temperature sterilization (60 ℃).
도 4는 활성탄 처리(0.2 g 활성탄/g 포도당) 후 저온 멸균시(60℃) 목질계 당화액으로부터 에탄올 발효능력(a: 포도당 농도, b: 에탄올 농도)을 나타낸 그래프이다.Figure 4 is a graph showing the ethanol fermentation capacity (a: glucose concentration, b: ethanol concentration) from the wood-based saccharification solution at low temperature sterilization (60 ℃) after activated carbon treatment (0.2 g activated carbon / g glucose).
도 5는 연속발효를 위한 내부여과 반응기 시스템의 모식도이다5 is a schematic diagram of an internal filtration reactor system for continuous fermentation.
도 6은 저온 멸균시(60℃) 목질계 당화액으로부터 내부여과 반응기를 이용한 연속 에탄올 발효능력을 나타낸 그래프이다.Figure 6 is a graph showing the continuous ethanol fermentation capacity using the internal filtration reactor from the saccharified liquor system at low temperature sterilization (60 ℃).
<도면의 주요부분에 대한 부호의 설명><Description of the symbols for the main parts of the drawings>
(1) : 내부여과기 모듈을 장착한 발효기(1): Fermenter equipped with internal filter module
(2) : 모터(2): motor
(3) : 압력게이지(3): Pressure gauge
(4) : 백플러싱(backflushing) 장치(4): backflushing device
(5) : 당화액 저장 탱크(5): saccharified liquid storage tank
(6) : 발효액 저장 탱크(6): fermentation broth storage tank
(7) : 잉여액 보관 용기(bleed chamber)(7): surplus liquid storage container (bleed chamber)
상기한 바와 같은 목적을 달성하고 종래의 결점을 제거하기 위한 본 발명의 실시예를 그 구성과 작용을 첨부도면에 연계시켜 상세히 설명하면 다음과 같다.Embodiments of the present invention for achieving the object as described above and to eliminate the drawbacks of the prior art will be described in detail in connection with the configuration and operation of the accompanying drawings.
본 발명은 폐목재, 도시유기성 폐기물을 포함한 목질계 바이오매스를 전처리·효소당화하여 당화액중의 포도당과 미생물을 반응시켜 에탄올을 생산하는 방법에 있어서,The present invention is a method for producing ethanol by reacting glucose and microorganisms in a saccharified solution by pretreatment and enzymatic saccharification of wood-based biomass including waste wood and municipal organic waste,
당화액의 멸균시 저온 멸균법을 사용하여 독성물질을 줄이는 단계를 거친 후당화액중의 포도당에 산업균주인 사카로마이세스 세레비제를 접종하여 에탄올을 발효 생산하는 회분식 방법과,Batch method of fermenting and producing ethanol by inoculating saccharomyces cerevisiae, an industrial strain, into glucose in the glycated saccharified solution after the step of reducing toxic substances by sterilizing the saccharified liquid,
또한 본 발명은 폐목재, 도시유기성 폐기물을 포함한 목질계 바이오매스를 전처리·효소당화하여 당화액중의 포도당과 미생물을 반응시켜 에탄올을 생산하는 방법에 있어서,The present invention also provides a method for producing ethanol by reacting glucose and microorganisms in saccharified solution by pretreatment and enzymatic saccharification of wood-based biomass including waste wood and municipal organic waste.
당화액의 멸균시 저온 멸균법과 활성탄 처리를 병행 사용하여 독성물질을 줄이는 단계를 거친 후 당화액중의 포도당에 산업균주인 사카로마이세스 세레비제를 접종하여 에탄올을 발효 생산하는 회분식 방법과,Batch method of fermenting and producing ethanol by inoculating saccharomyces cerevises, an industrial strain, into the glucose in the saccharified liquid after the step of reducing toxic substances by using low temperature sterilization method and activated carbon treatment at the time of sterilization of the saccharified liquid;
또한 본 발명은 폐목재, 도시유기성 폐기물을 포함한 목질계 바이오매스를 전처리·효소당화하여 당화액중의 포도당과 미생물을 반응시켜 에탄올을 생산하는 방법에 있어서,The present invention also provides a method for producing ethanol by reacting glucose and microorganisms in saccharified solution by pretreatment and enzymatic saccharification of wood-based biomass including waste wood and municipal organic waste.
당화액의 멸균시 저온 멸균법을 사용하여 독성물질을 줄이는 단계를 거친 후 당화액중의 포도당에 산업균주인 사카로마이세스 세레비제를 접종하여 내부여과기 모듈을 장착한 발효기를 이용하여 에탄올을 연속식으로 생산하는 방법으로 구성된다.After sterilization of the saccharified solution, the toxic substance is reduced by using low temperature sterilization method, and the glucose in the saccharified solution is inoculated with Saccharomyces cerevise, an industrial strain, and the ethanol is continuously fed using a fermenter equipped with an internal filter module. It consists of a method of production.
이하 본 발명을 실시예를 들어 상세히 기술하면 다음과 같다.Hereinafter, the present invention will be described in detail with reference to Examples.
실시예 1Example 1
: 저온 멸균법에 의한 목질계 바이오 매스로부터 고농도 에탄올 생산(회분식): Production of high concentration ethanol from woody biomass by low temperature sterilization (batch)
본 발명에 사용한 균주는 산업균주인 사카로마이세스 세레비제로 이 균주는 서영주정으로부터 분양받았다.The strain used in the present invention was Saccharomyces cerevisiae, an industrial strain, and the strain was distributed from Seoyoung.
접종을 위해 사용한 배지의 조성은 0.3% 이스트 익스트랙트(yeast extract), 0.3% 멀트 익스트랙트(malt extract), 0.5% 박토 펩톤(bacto peptone), 2% 포도당을 기본으로 하였으며 필요에 따라 농도를 조절하여 사용하였다.The composition of the medium used for inoculation was based on 0.3% yeast extract, 0.3% malt extract, 0.5% bacto peptone, and 2% glucose. Was used.
이때 사용한 목질계 당화액은 참나무를 우선 8L 증기폭쇄기를 이용하여 215℃에서 15분간 폭쇄하고, 30L 반응기를 이용하여 20 유닛(unit)의 필터페이퍼 활성도(filter paper activity)와 30유닛의 베타글루코시데이즈 활성도(β-glucosidase activity)를 가지게 조절한 셀룰라제(cellulase)와 베타글루코시데이즈 효소를 투여하여 50℃에서 3일간 당화한 후, 진공증발기를 이용하여 원하는 포도당 농도로 농축하여 사용하였다.The wood-based saccharification liquid used was first pulverized oak for 15 minutes at 215 ° C. using an 8L steam bomber, and 20 units of filter paper activity and 30 units of betaglucose using a 30L reactor. Cellulase and betaglucosidase enzyme adjusted to have a β-glucosidase activity were glycosylated at 50 ° C. for 3 days, and then concentrated to a desired glucose concentration using a vacuum evaporator.
목질계 당화 농축액을 이용한 에탄올 발효는 25ml의 농축액이 들어있는 100ml 플라스크를 이용하여 진탕배양기(shaking incubator)에서 발효를 수행하였다.Ethanol fermentation using wood-based saccharification concentrate was carried out in a shaking incubator using a 100 ml flask containing 25 ml of concentrate.
조업은 30℃, 150rpm에서 행하였으며 초기 pH는 5.0으로 일정하게 조절하였다.The operation was performed at 30 ° C. and 150 rpm and the initial pH was constantly adjusted to 5.0.
발효는 따로 영양배지를 첨가하지 않고 결과에 명시한 적절한 포도당 농도를 가지게 희석한 폭쇄재 당화액만을 사용하였다.The fermentation used only the explosive saccharification liquid diluted with the appropriate glucose concentration indicated in the result without the addition of nutrient medium.
도 1은 상기와 같이 준비한 당화액을 고온 멸균시(121℃) 목질계 당화액으로부터 에탄올 발효능력을 나타낸 그래프인데, 일반적인 고온 멸균을 행한 경우 목질계 당화액의 농도가 높아짐에 따라 발효속도가 늦어지고 접종량에 따라 원액을 포함하여 약 90g/L까지만 사용한 당을 모두 소모하였으며, 100g/L 이상에서는 발효가 수행되지 않았다.1 is a graph showing the ethanol fermentation capacity from the saccharified solution prepared at the high temperature sterilization (121 ℃) of the saccharified solution prepared as described above. The amount of sugar used up to about 90g / L, including the stock solution, was consumed according to the high inoculum dose, and fermentation was not performed at 100g / L or more.
이는 전처리 공정 중 생기는 발효저해물질 때문이다(참조: Palmqvist et al., Enzyme Microb. technol., 20: 286-293 (1997)).This is due to fermentation inhibitors during the pretreatment process (Palmqvist et al., Enzyme Microb. Technol., 20: 286-293 (1997)).
도 2는 당화액을 고온 멸균(121℃)후 활성탄 처리시 목질계 당화액으로부터 에탄올 발효능력을 나타낸 그래프인데, 저해물질의 영향을 감소시키기 위해 자주 사용되는 활성탄을 처리해준 경우 어느 정도 효과가 있었으나 목질계 당화액의 포도당 농도 100 g/L이상에서는 역시 발효가 이루어지지 않았다.Figure 2 is a graph showing the ethanol fermentation capacity from the wood-based saccharification solution when the saccharified solution at high temperature sterilization (121 ℃) activated carbon treatment, there was some effect when treated with activated carbon which is often used to reduce the effects of inhibitors Fermentation did not occur at the glucose concentration of 100 g / L or more of the wood-based saccharified solution.
그러나 도 3 (a, b)에서 당화액을 저온 멸균(60∼70℃, 30∼120분)을 하고 발효를 수행한 결과 고온 멸균(121℃, 15분)시 발효가 이루어지지 않은 100g/L 기질뿐만 아니라 140 g/L까지도 정상적 발효가 가능함을 보여주었다.However, as a result of the sterilization of the saccharified solution at low temperature sterilization (60-70 ° C., 30-120 minutes) in FIG. 3 (a, b), the fermentation was not performed at high temperature sterilization (121 ° C., 15 minutes). It was shown that normal fermentation was possible up to 140 g / L as well as substrate.
즉, 도 3 (a, b)는 저온 멸균시(60 ∼ 70℃, 30 ∼ 120분) 목질계 당화액으로부터 에탄올 발효능력(a: 포도당 농도, b: 에탄올 농도)을 나타낸 그래프인데, 특히 140 g/L 기질사용시 사카로마이세스 세레비제를 20%(v/v) 접종하여준 경우 발효가 2일 후 거의 완료됨을 알 수 있었으며, 이때 발효생성된 에탄올 농도와 생산성은 각각 67g/L와 1.0 g/L/h이었다.That is, Figure 3 (a, b) is a graph showing the ethanol fermentation capacity (a: glucose concentration, b: ethanol concentration) from the wood-based saccharification liquid at low temperature sterilization (60 ~ 70 ℃, 30 to 120 minutes), especially 140 Fermentation was almost complete after 2 days when inoculated with 20% (v / v) of Saccharomyces cerevises when using g / L substrate, and the ethanol concentration and productivity of fermentation were 67g / L and 1.0, respectively. g / L / h.
도 4 는 저온 멸균후(60 ∼ 70℃) 활성탄 처리(0.2 g 활성탄/g 포도당)시 목질계 당화액으로부터 에탄올 발효능력(a: 포도당 농도, b: 에탄올 농도)을 나타낸그래프인데, 기질농도 170 g/L를 사용할 때에도 100시간 이상의 적응기(lag period)가 존재하였지만 발효가 이루어져 기존의 고온 멸균법에 비해 발효가 효과적이었으며 활성탄 처리를 추가하면 더욱 효과적이었다.Figure 4 is a graph showing the ethanol fermentation capacity (a: glucose concentration, b: ethanol concentration) from the wood-based saccharification when activated carbon treatment (0.2 g activated carbon / g glucose) after low temperature sterilization (60 ~ 70 ℃), substrate concentration 170 Even when g / L was used, a lag period of 100 hours or more existed, but the fermentation was more effective than the conventional high temperature sterilization method, and the addition of activated carbon treatment was more effective.
실시예 2Example 2
: 저온 멸균법을 이용한 내부여과기 모듈을 장착한 발효기에서의 고농도, 고생산성 에탄올 생산 방법(연속식): High concentration and high productivity ethanol production method in fermenter equipped with internal filter module using low temperature sterilization method (continuous type)
도 5는 연속발효를 위한 내부여과기 모듈을 장착한 발효기 시스템의 모식도인데, 동 실시예 2에서는 실시예 1에 예시한 회분식과 비교하여 저온 멸균법을 이용하여 내부여과기 모듈을 장착한 발효기에서 연속식 발효를 수행하여 에탄올의 생산성 향상효과를 관찰하였다.FIG. 5 is a schematic diagram of a fermenter system equipped with an internal filter module for continuous fermentation. In Example 2, continuous fermentation is performed in a fermenter equipped with an internal filter module using low temperature sterilization as compared to the batch illustrated in Example 1. Was carried out to observe the effect of improving the productivity of ethanol.
사용한 3L 발효기(1)의 발효액의 부피는 1.5L이고, 연속식 발효시 발효기에 공급할 새로운 배지는 포도당 100 ∼ 180 g/L가 포함된 목질계 당화액 만을 사용하였다.The volume of the fermentation broth of the used 3L fermenter (1) is 1.5L, and only the wood-based saccharification solution containing 100-180 g / L of glucose was used as a new medium to be supplied to the fermentor during continuous fermentation.
발효기(1) 내부에 장착된 여과기 모듈은 내부반경, 외부반경, 높이가 각각 8, 11, 80 mm인 13개의 원통형 세라믹 튜브로 구성되어 있어, 여과 표면적이 360 cm2이었다.The filter module mounted inside the fermenter 1 is composed of 13 cylindrical ceramic tubes having an inner radius, an outer radius, and a height of 8, 11 and 80 mm, respectively, and has a filtration surface area of 360 cm 2 .
이때 사용한 여과기의 재질은 세라믹이었으며, 기공크기(pore size)는 0.3㎛이하로 구성되어 있고 여과 표면적이 발효액 내부에 배치된다.The material of the filter used was ceramic, the pore size (pore size) is composed of less than 0.3㎛ and the filtration surface area is disposed inside the fermentation broth.
부호 (2)는 발효기를 교반하는 모터, (3)은 여과기 모듈을 빠져 나오는 액체의 부압을 측정하는 압력게이지, (4)는 여과기 모듈이 폐색될 시 여과층을 거꾸로 불어내는 백플러싱(backflushing) 장치, (5)는 당화액 저장 탱크, (6)은 생산물인 발효액 저장탱크, (7)은 백플러싱 등을 실시할 때 발생하는 잉여액 보관 용기이다.Symbol (2) is a motor for stirring the fermenter, (3) is a pressure gauge for measuring the negative pressure of the liquid exiting the filter module, (4) backflushing which blows the filter layer upside down when the filter module is blocked. The apparatus (5) is a saccharification liquid storage tank, (6) is a fermentation liquid storage tank which is a product, and (7) is a surplus liquid storage container which arises when backflushing etc. are performed.
당화액 저장 탱크(5)에 보관된 고농도 당화액(100g/L∼180g/L)은 펌프를 통하여 발효기(1)로 투입되고 모터(2)로 교반되면서 사카로마이세스 세레비제에 의하여 에탄올로 발효된다. 동시에 발효기(1) 내부의 액은 여과기 모듈을 통하여 발효액 저장탱크(6)로 연속적으로 빠져 나오며 내부에 자라난 사카로마이세스 세레비제 균체와 고형물은 여과기 모듈에서 여과된다.The high concentration of saccharified liquid (100 g / L to 180 g / L) stored in the saccharified liquid storage tank (5) was introduced into the fermenter (1) through a pump and stirred by a motor (2) to ethanol by Saccharomyces cerevisiae. Fermented. At the same time, the liquid in the fermenter 1 is continuously discharged into the fermentation broth 6 through the filter module, and the Saccharomyces cerevisiae cells and solids grown therein are filtered in the filter module.
이와 같이 연속발효를 계속하면 여과기 모듈에 고형물이 침적되어 압력게이지(3)가 마이너스로 떨어지며 발효액의 배출이 힘들어지게 되는데 주기적으로 백플러싱(backflushing) 장치(4)를 가동하여 여과층을 역세함으로서 발효액의 배출이 용이하도록 한다.If the continuous fermentation is continued, the solids are deposited on the filter module, so that the pressure gauge (3) falls to the negative and the discharge of the fermentation broth becomes difficult. The fermentation broth is backwashed by periodically operating the backflushing device (4). Should be easy to discharge.
연속식 발효는 먼저 회분식으로 발효를 수행한 뒤에 기질이 전부 소모된 후 희석속도(dilution rate) 0.22 h-1이상으로 운전하였다.Continuous fermentation was first performed in a batch fermentation, and then the substrate was consumed and operated at a dilution rate of 0.22 h −1 or more.
발효는 30 ℃에서 수행하였으며, 발효기내의 세포의 활성을 높게 유지하기 위해 0.5 ∼ 1.0vvm으로 공기를 공급하며 발효액중의 효모 균체를 내부여과 하였다.The fermentation was carried out at 30 ℃, in order to maintain high activity of the cells in the fermenter was supplied with air at 0.5 ~ 1.0vvm and yeast cells in the fermentation broth were internal filtration.
도 6은 저온 멸균시(60℃) 목질계 당화액으로부터 내부여과기 모듈을 장착한발효기를 이용하여 에탄올 발효능력을 나타낸 그래프인데, 발효기를 이용하여 저온 멸균한 목질계 당화액으로부터 연속 발효를 수행한 결과 포도당 137g/L과 180 g/L가 포함된 당화액의 경우 세포 농도와 에탄올 농도가 각각 1.4 ×109cells/ml, 1.5 ×109cells/ml와 58.8 g/L, 76.9 g/L에 이르러, 기존 방법에 비교하여 매우 높은 세포 농도와 에탄올 농도를 얻을 수 있었다.6 is a graph showing the ethanol fermentation capacity using the fermenter equipped with an internal filter module from the wood-based saccharified solution at low temperature sterilization (60 ℃), a continuous fermentation from the wood-based saccharified solution sterilized by using a fermenter Results For saccharified solutions containing 137 g / L and 180 g / L of glucose, the cell concentration and ethanol concentration were 1.4 × 10 9 cells / ml, 1.5 × 10 9 cells / ml, 58.8 g / L, and 76.9 g / L, respectively. As a result, very high cell concentration and ethanol concentration were obtained compared to the conventional method.
이때 에탄올 생산 수율은 0.43 g 에탄올/g 포도당이었다.Ethanol production yield was 0.43 g ethanol / g glucose.
또한 이때의 에탄올 생산성은 최고 16.9 g/L/h로 실시예 1에 예시한 회분식 배양에서의 값(1.0 g/L/h)나 고온 멸균에 의한 발효기에서의 값(6.7 g/L/h, 참조: Lee et al., Korean J. of Chem, Eng., 13: 453-456 (1996))보다 월등히 높은 결과이다.In addition, the ethanol productivity at this time is up to 16.9 g / L / h, the value in the batch culture illustrated in Example 1 (1.0 g / L / h) or the value in the fermenter by high temperature sterilization (6.7 g / L / h, See: Lee et al., Korean J. of Chem, Eng., 13: 453-456 (1996)).
저온 멸균법을 이용한 회분식 배양에서처럼, 내부여과기 모듈을 장착한 발효기를 이용한 연속 배양에서도 저온 멸균에 의한 오염문제는 발생하지 않아 목질계 당화액을 이용한 에탄올 등 유용물질 생산에 있어서 매우 효율적인 공정이었으며, 이를 폐목재, 도시유기성 폐기물로부터 에탄올 생산에 응용하는 경우 생산성이 높으므로 경제적인 측면에서 매우 긍정적인 결과를 얻었다.As in batch cultivation using low temperature sterilization method, the contamination problem by low temperature sterilization did not occur even in continuous culture using a fermenter equipped with an internal filter module, which was a very efficient process for producing useful substances such as ethanol using wood-based saccharification liquid. The applications for ethanol production from wood and municipal organic wastes have been highly economical, with high productivity.
본 발명은 상술한 특정의 바람직한 실시예에 한정되지 아니하며, 청구범위에서 청구하는 본 발명의 요지를 벗어남이 없이 당해 발명이 속하는 기술분야에서 통상의 지식을 가진 자라면 누구든지 다양한 변형실시가 가능한 것은 물론이고, 그와같은 변경은 청구범위 기재의 범위 내에 있게 된다.The present invention is not limited to the above-described specific preferred embodiments, and various modifications can be made by any person having ordinary skill in the art without departing from the gist of the present invention claimed in the claims. Of course, such changes will fall within the scope of the claims.
이상에서 상세히 설명하였듯이, 본 발명은 저온 멸균을 이용하여 목질계 바이오 매스로부터 고농도 에탄올을 생산하는 방법을 제공한다.As described in detail above, the present invention provides a method for producing high concentration ethanol from woody biomass using low temperature sterilization.
또한 본 발명에 의하면, 기존의 고온 멸균법을 사용하여서는 발효가 불가능한 180g/L 포도당 농도의 고농도 목질계 당화액을 저온 멸균법과 내부여과기를 장착한 연속 발효기를 이용하여 기존 방법으로는 얻을 수 없는 76.9g/L의 에탄올 농도와 16.9 g/L/h의 에탄올 생산성을 얻을 수 있었다.In addition, according to the present invention, the high-density wood-based saccharification solution of 180g / L glucose concentration which cannot be fermented using the existing high-temperature sterilization method can not be obtained by the conventional method using a continuous fermenter equipped with low-temperature sterilization method and an internal filter 76.9g Ethanol concentration of / L and ethanol productivity of 16.9 g / L / h were obtained.
따라서 본 발명은 미생물을 이용하여 목질계 바이오매스로부터 유용한 물질을 생산하기 위해 필수적인 목질계 바이오매스의 전처리, 당화 및 멸균시 생기는 독성물질의 영향을 저온 멸균과 고생산성 발효로 최소화하여 유용물질 생성률을 향상시켜 값싸고 풍부한 목질계 바이오매스로부터 미생물을 이용하여 에탄올 등을 포함한 유용물질을 생산하는 공정의 산업화에 기여할 것이다.Therefore, the present invention minimizes the effects of toxic substances generated during pretreatment, saccharification and sterilization of wood based biomass, which are essential for producing useful materials from wood based biomass using microorganisms, by minimizing low temperature sterilization and high productivity fermentation. It will contribute to the industrialization of the process of producing useful substances, including ethanol, using microorganisms from cheap and abundant woody biomass.
Claims (7)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1019990061537A KR100318755B1 (en) | 1999-12-24 | 1999-12-24 | Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1019990061537A KR100318755B1 (en) | 1999-12-24 | 1999-12-24 | Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20010058065A KR20010058065A (en) | 2001-07-05 |
| KR100318755B1 true KR100318755B1 (en) | 2001-12-28 |
Family
ID=19629144
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1019990061537A KR100318755B1 (en) | 1999-12-24 | 1999-12-24 | Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100318755B1 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012087068A2 (en) * | 2010-12-24 | 2012-06-28 | 한국화학연구원 | Treatment method for biomass to maximize sugar yield, and additive used in same |
| KR101390254B1 (en) * | 2010-12-24 | 2014-05-02 | 한국화학연구원 | Treatment method of biomass to maximize sugar yield and additive used therein |
| WO2015026138A1 (en) * | 2013-08-22 | 2015-02-26 | 전남대학교산학협력단 | Method for removing limonene and device for removing limonene in biomass saccharification liquid, and bioethanol production method using the removal method |
| KR101541762B1 (en) * | 2013-11-11 | 2015-08-05 | 한국생산기술연구원 | A method for producing fermented sugar and alcohol from mass-producing bread waste |
| US9657318B2 (en) | 2013-04-04 | 2017-05-23 | Korea Institute Of Science And Technology | Electrochemical detoxification method of wood-based hydrolysate for producing biochemicals or biofuels |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040029658A (en) * | 2002-10-02 | 2004-04-08 | 류하수 | Ethanol continuous fermentation system |
| KR100965851B1 (en) * | 2007-10-11 | 2010-06-28 | 전남대학교산학협력단 | Processes for the pretreatment of lignocellulosic biomasses by popping method, and processes for the production of saccharides and bio-ethanol using the same |
| KR101035510B1 (en) * | 2008-10-28 | 2011-05-20 | 현대제철 주식회사 | Cooler for guide roller in hot rolling facilities |
| KR101711571B1 (en) | 2015-05-29 | 2017-03-02 | 신호테크 주식회사 | Environment Friendly and Fire-Retardant Material EVA Panel |
-
1999
- 1999-12-24 KR KR1019990061537A patent/KR100318755B1/en active IP Right Grant
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012087068A2 (en) * | 2010-12-24 | 2012-06-28 | 한국화학연구원 | Treatment method for biomass to maximize sugar yield, and additive used in same |
| WO2012087068A3 (en) * | 2010-12-24 | 2012-08-16 | 한국화학연구원 | Treatment method for biomass to maximize sugar yield, and additive used in same |
| KR101390254B1 (en) * | 2010-12-24 | 2014-05-02 | 한국화학연구원 | Treatment method of biomass to maximize sugar yield and additive used therein |
| US10081823B2 (en) | 2010-12-24 | 2018-09-25 | Korea Research Institute Of Chemical Technology | Treatment method for biomass to maximize sugar yield, and additive used in same |
| US9657318B2 (en) | 2013-04-04 | 2017-05-23 | Korea Institute Of Science And Technology | Electrochemical detoxification method of wood-based hydrolysate for producing biochemicals or biofuels |
| WO2015026138A1 (en) * | 2013-08-22 | 2015-02-26 | 전남대학교산학협력단 | Method for removing limonene and device for removing limonene in biomass saccharification liquid, and bioethanol production method using the removal method |
| KR101541762B1 (en) * | 2013-11-11 | 2015-08-05 | 한국생산기술연구원 | A method for producing fermented sugar and alcohol from mass-producing bread waste |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20010058065A (en) | 2001-07-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Velásquez-Riaño et al. | Production of bacterial cellulose from alternative low-cost substrates | |
| JP5149785B2 (en) | Treatment of biomass to obtain ethanol | |
| US6835560B2 (en) | Process for ozonating and converting organic materials into useful products | |
| JP4038577B2 (en) | Alcohol production system and alcohol production method | |
| JP2011514806A (en) | A method for the conversion of plant materials into fuels and chemicals by the continuous action of two microorganisms | |
| AU2009262334A1 (en) | Method of producing yeast biomass | |
| KR100318755B1 (en) | Process for producing ethanol with high concentration from wood hydrolysate using low-temperature sterilization | |
| Perego et al. | Experimental study of hydrogen kinetics from agroindustrial by-product: optimal conditions for production and fuel cell feeding | |
| CN108504584A (en) | A kind of culture medium and application for being suitable for improving co-fermentation of glucose and xylose saccharomyces cerevisiae and being resistant to a variety of pretreatment mortifiers | |
| US4351905A (en) | Horizontal fermenter | |
| Jin et al. | A bioprocessing mode for simultaneous fungal biomass protein production and wastewater treatment using an external air‐lift bioreactor | |
| JP2008259517A (en) | Equipment and method for producing lactic acid from cellulose | |
| JP4170016B2 (en) | Lactic acid production apparatus and method for producing lactic acid from cellulose | |
| Lee et al. | A cell retention internal filter reactor for ethanol production using tapioca hydrolysates | |
| Kim et al. | Continuous production of citric acid from dairy wastewater using immobilized Aspergillus niger ATCC 9142 | |
| Lee et al. | Continuous ethanol production from wood hydrolysate by chemostat and total cell retention culture | |
| CN106282154B (en) | A kind of preparation method and applications with the co-immobilization mycelium pellet for removing multiple pollutant function | |
| JP3004509B2 (en) | Method and apparatus for producing ethanol from microalgae | |
| CN113025497B (en) | Bacterial strain for efficiently degrading anthraquinone dyes and method for improving decoloring efficiency by using traditional Chinese medicine waste residues | |
| Zohri et al. | Continuous Ethanol Production from Molasses via Immobilized Saccharomyces cerevisiae on Different Carriers on Pilot Scale | |
| JPH0731484A (en) | Method for biologically producing hydrogen | |
| CN112979355B (en) | Method for treating kitchen waste by self-made composite hydrolase | |
| CN218345419U (en) | Continuous fermentation device | |
| Taniguchi et al. | Continuous ethanol production by cell-holding culture of yeasts | |
| RU2404247C2 (en) | Method of obtaining butanol |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20121213 Year of fee payment: 12 |
|
| FPAY | Annual fee payment |
Payment date: 20131205 Year of fee payment: 13 |
|
| FPAY | Annual fee payment |
Payment date: 20141211 Year of fee payment: 14 |
|
| FPAY | Annual fee payment |
Payment date: 20151105 Year of fee payment: 15 |
|
| FPAY | Annual fee payment |
Payment date: 20170112 Year of fee payment: 16 |
|
| FPAY | Annual fee payment |
Payment date: 20171106 Year of fee payment: 17 |
|
| FPAY | Annual fee payment |
Payment date: 20181105 Year of fee payment: 18 |
|
| FPAY | Annual fee payment |
Payment date: 20191105 Year of fee payment: 19 |