TWI546379B - Microrganism for forming fermentation products through fermentation of sugars - Google Patents

Description

Microorganism for fermenting sugar to form a fermentation product

The present invention relates to a microorganism for fermentation, and more particularly to a microorganism capable of transforming a plastid containing a sequence of a mutated enzyme into a microorganism, thereby fixing carbon dioxide and improving fermentation efficiency.

All along, the production of fermented foods relies on microorganisms. Microorganisms are connected to raw materials. During the growth process, the required enzymes are produced, and the process of decomposing organic matter into beneficial substances in human life is called fermentation; therefore, this metabolic activity is actually catalyzed. The performer is an enzyme produced by microorganisms.

In the process of microbial fermentation, it can be divided into aerobic fermentation and anaerobic fermentation depending on whether oxygen is required. For example, mold culture and acetic acid fermentation are aerobic fermentation; alcohol fermentation and lactic acid fermentation are anaerobic fermentation. According to different raw material types and devices, it can be divided into solid fermentation and liquid fermentation, solid fermentation such as rice and sorghum fermentation. Because of the low water content of raw materials, the tolerance of mixed bacteria is high, and it is suitable for the cultivation of open space. For example, mold culture; liquid fermentation, because of high water content, rapid growth of microorganisms, easy to contaminate, it is often used in closed culture, such as the fermentation process requires oxygen, and then into the sterile air.

In recent years, in order to utilize the principle of microbial fermentation, it is necessary to prepare human daily life. Fermented products have gradually developed the industry of fermentation engineering. Fermentation engineering is an area that solves the engineering problems of industrial production by fermentation process. Fermentation engineering divides the fermentation industrial process that realizes the fermentation process into three stages: strain, fermentation and refining (including wastewater treatment) from the engineering point of view. These three stages have their own engineering problems, which are generally called respectively. It is the upstream, midstream and downstream projects of the fermentation project. The three stages of the fermentation project have their respective process principles and equipment and process control principles, which together constitute the principle of fermentation engineering.

Among them, the fermentation engineering involves the chemistry of carbon, and the carbon enters the ecological circle in the form of carbon dioxide in the metabolic process of fermentation engineering. The ribulose bisphosphate carboxylase/oxygenase (RubisCO) is an enzyme that plays a role in converting carbon dioxide into biomass (biomass) in nature. This is a common enzyme; however, ribulose bisphosphate carboxylase/oxygenase is currently a slower metabolic enzyme (Kcat~3/s) in bioengineering. It is not clear why its rate is so slow. And low specificity, and many

However, in this fermentation process, the use of microorganisms to produce small molecule chemicals such as bioethanol or bio-butanol is focused on how to effectively produce it, and after taking into account the production of small molecule chemicals, once mass production is started, The result is a large amount of carbon dioxide emissions, such as the production of a molar ethanol will produce a mole of carbon dioxide, few people focus on the fixation or recycling of carbon dioxide and its further use. Only in U.S. Patent No. 20080085341, which provides a method for forming a fermentation product using at least one heterologous gene sequence which exhibits an exogenous protein capable of converting a foreign carbon source into and fixing carbon dioxide. To the desired fermentation product; although it has been mentioned that immobilized carbon dioxide makes the environment oxidize The amount of carbon is no longer increasing, but it does not further improve industrial applications such as fermentation efficiency, capacity, and bioengineering.

In view of the above, it is an object of the present invention to provide a microorganism which can focus on the fixation and recovery of carbon dioxide during the fermentation process, thereby reducing the amount of carbon dioxide emissions in the fermentation industry and reducing the burden of excessive carbon dioxide on the earth.

At the same time, another object of the present invention is to transform a plastid containing a sequence of a mutant enzyme into a microorganism to enhance the efficiency of fermentation under the premise of fixing carbon dioxide.

In order to achieve the above object, the present invention provides a microorganism for forming a fermentation product by fermentation, which comprises a plastid or a chromosome containing a mutated enzyme sequence, and the mutated or non-mutated enzyme comprises a structure selected from the following sequence a group of plastids or chromosomes: pACYC184-PrkA/pET-rbcLSm1/tktA (JB/PTA) and pACYC184-PrkA/pET-rbcLSm1/talB (JB/pTB); wherein the receptor is a six-carbon sugar, A hexasaccharide derivative or a combination thereof; wherein the carbon dioxide emission per unit C2 organic matter consumption after fermentation of the plastid or chromosome having the sequence structure is reduced by 6.25%-15.6% compared with the wild type strain.

In a preferred embodiment of the invention, the microorganism can be a fungus.

Preferably, the fungus can be a genus of yeast.

In another preferred embodiment of the invention, the microorganism can be a bacterium or an archaea or a combination thereof.

Preferably, the bacteria may be Escherichia coli, Clostridium acetobutylicum, Bacillus subtilis (Bacillus). Subtilis), Pseudomonas putia, cyanobacteria, Cupriavidus spp., or Zymomonas mobilis, or a combination thereof.

In another preferred embodiment of the invention, the microorganism can be an alga.

Preferably, the hexasaccharide derivative is a disaccharide, oligosaccharide or polysaccharide polymerized by a hexose.

Many of the features and advantages of the present invention will become more apparent from the detailed description of the appended claims.

The first plot shows the growth curves of various strains only under aerobic conditions in LB medium.

The second graph shows the (A) growth curve and (B) sugar content of various strains in aerobic conditions of LB medium containing arabinose.

The third figure is a schematic diagram showing the ratio of carbon dioxide produced by fermentation of Escherichia coli to acetic acid, carbon dioxide and ethanol.

The fourth graph is a bar graph comparing the carbon dioxide emissions per unit of sugar consumption in each strain of the fermentation tank.

The fifth graph is a bar graph comparing the yield of ethanol and acetic acid per unit sugar consumption in each strain of the fermentation tank.

The sixth graph is a bar graph comparing the CO2 emissions per unit of ethanol and acetic acid production in each strain of the fermentation tank.

The detailed description of the present invention is intended to be illustrative of the embodiments of the invention, and is not intended to represent the invention. real The order of the functions and steps mentioned in the examples is used to construct and operate the embodiments. However, the same or equivalent functions or sequences may be achieved by different embodiments.

For the sake of convenience, the embodiments employed in the specification, examples, and appended claims are hereby incorporated by reference. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art.

The singular forms "a", "the", and "the"

The term "fixed" as used herein refers to the conversion of carbon dioxide and/or the incorporation of carbon atoms of carbon dioxide into organic molecules such that the carbon dioxide produced in the fermentation process can be recycled indefinitely.

The term "microorganism" is defined herein as an organism that can form a fermentation product for fermentation by fermentation, and is not limited to conventional microorganisms.

The present invention provides a microorganism for forming a fermentation product by fermentation, which comprises a plastid or a chromosome containing a mutated or non-mutated enzyme sequence, which may be a nucleotide sequence or a protein sequence, which is mutated or The non-mutated enzyme is selected from the group consisting of the following EC number (Enzyme Commission number): 2.7.1, 5.1.3, 5.3.1, 4.1.1, 2.2.1, and 3.1.3; The receptor is a six carbon sugar, a six carbon sugar derivative or a combination thereof.

Among them, EC number 2.7.1 is preferably phosphoribulokinase (EC number: 2.7.1.19); EC number 5.1.3 is preferably ribulose-phosphate 3-epimerase (Ribulose-phosphate 3-epimerase, EC number: 5.1.3.1), 5-phosphate ribulose 4-isomerase (L-ribulose-5-phosphate 4-epimerase, EC number: 5.1.3.4), L-ribulose-5-phosphate 3-epimerase, EC number: 5.1 .3.22); EC number 5.3.1 is preferably ribose 5-phosphate isomerase/phosphopentose epimerase (EC number: 5.3.1.6); EC number 4.1.1 Preferably, ribulose-1, 5-bisphosphate carboxylase/oxygenase (RubisCo), EC number: 4.1.1.39); EC number 2.2.1 is preferably transaldolase (transaldolase, EC number: 2.2.1.2), acetoin-ribose-5-phosphate transaldolase (EC number: 2.2.1.4), fluorohydroxybutyrate transaldehyde Alcoholase (EC number: 2.2.1.8), transketolase (EC number: 2.2.1.1), formaldehyde transketolase (EC number: 2.2.1.3); EC number 3.1. 3 is preferably sedoheptulose 1,7-bisphosphatase (EC number: 3.1.3.37).

The fermentation product may be hydrogen, nucleotides, amino acids, alcohols such as ethanol, butanol, isobutanol, propylene glycol isomers, sorbitol, alkenes such as ethylene, propylene, isoprene. Isoprene, organic acids such as fatty acids, succinic acid, shikimic acid, acetic acid, esters such as biodiesel, poly(3-hydroxybutyrate, PHB), alkene Hydrocarbons, organic acids such as succinic acid, itaconic acid, levulinic acid, 3-hydroxypropiohic acid, polylactic acid fiber (polylactic Acid) and biomass (biomass).

Among them, poly(3-hydroxybutyrate) is a biodegradable material

Among them, the microorganism of the present invention can be used for fixing carbon dioxide.

In a preferred embodiment of the invention, the microorganism can be a fungus (Fungi).

Among them, the above fungus may be a genus of the genus Saccharomycesspp.

In another preferred embodiment of the invention, the microorganism can be a bacterium.

Among them, the above bacteria may be Escherichia coli., Clostridium acetobutylicum, Bacillus subtilis, Pseudomonas putia, cyanobacteria, Chromium (Cupriavidus spp.) or Zymomonas mobilis or a combination thereof.

In another preferred embodiment of the invention, the microorganism can be an algae.

Among them, the derivative of the hexasaccharide may be a disaccharide, oligosaccharide or polysaccharide polymerized by a hexose. It should be noted that the disaccharide, oligosaccharide or polysaccharide comprises, but is not limited to, disaccharides, oligosaccharides or polysaccharides polymerized by hexoses known to those of ordinary skill in the art.

In addition, the present invention can continuously fix the carbon dioxide generated after the fermentation, and the gas of the fermentation product only leaves hydrogen gas. Therefore, the present invention can also improve the efficiency of hydrogen removal and reduce the cost of hydrogen purification.

The examples described below are provided to illustrate certain aspects of the invention and to assist those skilled in the art to practice the invention. These examples are not to be construed as limiting the scope of the invention.

Instance

The present invention will further describe the nature and application of the modified microorganisms in the fermentation with reference to the following examples, which are intended to be illustrative only and not intended to limit the scope of the invention.

Instance

The present invention will further describe the nature and application of the modified microorganisms in the fermentation with reference to the following examples, which are intended to be illustrative only and not intended to limit the scope of the invention.

Example 1

The strain used in the fermentation process in this experiment is E.coli BL21(DE3) (hereinafter referred to as Escherichia coli), and the enzyme to be transferred into Escherichia coli is the mutated ribulose bisphosphate carboxylase/oxygenase. (RbcLS) and ribulose phosphate kinase (PrkA). The allele mutation sequence of the two enzymes is described in Parikh et. al., Directed evolution of RuBisCO hypermorphs through genetic selection in engineered E.coli, 2006, Protein Engineering, Design & Selection, 19:113. In the method of treatment, the mutated sequence itself, its N-terminal coding, restriction enzymes and the like, and the subclone method, and the treatment method of the strain of the present invention are the same as the paper, which is a common knowledge in the field. The known techniques are not described here. After treatment with each step, the final structure of the treated plasmid was pACYC184-PrkA, pET-rbcLS, pET-rbcLSm1, and transformed into E. coli and named J1, J2 and J3, respectively. In addition, pACYC184-PrkA and pET-rbcLS were additionally transformed into Escherichia coli, which is called JA; pACYC184-PrkA and pET were also added. -rbcLSm1 is co-transformed into E. coli and is referred to as JB. In addition, although the present embodiment inserts the condition of the desired sequence into the plastid, it is merely exemplary and not limiting. For example, the present invention can also insert the condition of the desired sequence into the chromosome and insert the chromosome. The advantage is that the bioengineering system of the invention presents a more stable state. Furthermore, this embodiment uses the T7 startup subsystem for testing, but it is not limiting, and any conventional startup subsystem can be applied to the present invention. It should be noted that this example only uses the above-mentioned mutant enzyme as an experimental description, and other related mutant-treated enzymes, such as: ribulose bisphosphate carboxylase activating enzyme, phosphoribril kinase, 1 , 5-ribulose ribulose epimerase, ribulose bisphosphate carboxylase/oxygenase type I, ribulose bisphosphate carboxylase/oxygenase type II, ribulose diphosphate Carboxylase/oxygenase type III, ribulose bisphosphate carboxylase/oxygenase type IV, or an enzyme that metabolizes sugar-related substances, such as: pentose phosphate isomerase, 5-phosphate Ribosomerase, transaldolase, transketolase and sedoheptulose-1,7-bisphosphatase can be used in this experiment.

Example 2

The culture medium used for the activation of the strain is a sterile protein medium (Luria Broth Medium) (hereinafter referred to as LB medium), and the composition thereof is NaCl 10 g/L; tryptone 10 g/L; and yeast extract. (Yeast Extract) 5g/L. Wherein the wild-type E. coli by adding antibiotics, as a system for controlling group; and applying the antibiotic chloramphenicol as J1 (Chloramphenicol) 34 μ g; J2 and J3 are Shijia Kang neomycin (Kanamycin) 50 μ g ; Finally, JA and JB are respectively applied to both chloramphenicol and 34 μ g amphotericin Kang 50 μ g. Isopropyl-β-D-thiogalactoside (IPTG) was selectively added during the cultivation according to the needs of the control group or the experimental group.

After that, the batch culture procedure was carried out, and Escherichia coli was cultured in a 125 mL sterile grooved conical flask. The Erlenmeyer flask contained 25 mL of LB medium, and the culture environment was activated in an aerobic environment at 37 ° C, 200 r. pm for 12 hours (preculture) ) to increase the activity of the cells and to take samples at appropriate times.

Next, the fermentation tank is subjected to semi-anaerobic fermentation in a 3L tank, and 1L LB medium is in the tank; the culture environment is 37 ° C, 150 r. pm, and the amount of carbon dioxide dissolved in the LB medium is continuously observed, and then taken at intervals. , metabolite analysis and sugar content in LB medium.

Wherein, the LB medium used contains 20 g/L of arabinose (L-Arabinose) and the corresponding antibiotic; however, the arabinose is merely exemplary in the embodiment, not limiting, and other five carbon sugars, For example, xylose or a combination with other five carbon sugars, as well as six carbon sugars such as glucose, galactose, fructose, mannose or combinations thereof, can be readily utilized in this experiment.

Example 3

After the activation step is completed, the cell density of Escherichia coli is measured by a spectrophotometer, and the amount of bacteria in the main culture is determined by the value of OD _600. This step mainly uses the absorbance of Escherichia coli to measure the concentration of the E. coli culture solution, thereby The growth of E. coli is estimated, so it is usually used to refer to the cell density of the cells. Before the measurement, warm up for 10 minutes, then adjust the wavelength to 600nm to measure the absorption value of each sample; firstly, put the blank sample into the zero adjustment operation, if the absorption value is greater than 0.8, dilute to the appropriate magnification, and then measure the absorption. Value; at the beginning of the culture, the initial OD ₆₀₀ value of each of the cells was about 0.050.

Example 4

A quantitative test of arabinose was carried out during fermentation to compare the fermentation efficiency. First, take a 1m sample and centrifuge at 13,300rpm for 10 minutes. Then remove the supernatant and dilute to the appropriate ratio. Add DNS reagent (10g/L dinitrosalicylic acid, 16g/) in a ratio of 1:1. L sodium hydroxide and 300 g/L potassium sodium tartrate). Then, the mixture was heated in a water bath at 100 ° C for 10 minutes in the dark, and then cooled in the dark for 5 minutes. Finally, the absorption value of the wavelength at 550 nm is measured by a spectrophotometer. If the absorption value is greater than 0.8, the dilution is performed to an appropriate magnification, and the absorption value is measured.

The content of reducing sugar in the fermentation broth can be known from the previous calibration curve.

Example 5

Next, E. coli was separately placed into the mutated ribulose ribose carboxylase/oxygenase (RbcLS mutant A), ribulose kinase (PrkA), and transaldolase B (transaldolase B). Or a transketolase A gene fragment. Since transaldolase and transketolase are the most important enzymes in the non-oxidative pentose phosphate pathway, this pathway is used to convert six-carbon sugar into five-carbon sugar for metabolism. Transaldolase and transketolase play the role of converting two carbons and three carbons, respectively.

First, first remove some bacteria liquid into the colorimetric cell, and put it into UV/Vis, set the wavelength to 600nm and measure its absorbance, which is the cell concentration.

Next, the measurement of the reducing sugar was carried out. First, the method of preparing the calibration curve by the absorption value v.s. concentration is similar to that of the example 4, and details are not described herein again.

After preparing the calibration curve, 1 ml of the sample was mixed with +1 ml of DNS reagent (1:1). Take 1 ml of deionized water and 1 ml of DNS reagent as a blank test group (Blank). then Heat in a 100 ° C water bath for 10 minutes and cool in the dark for 5 minutes. Next, after setting the wavelength of the spectrophotometer to 550 nm, the blank test group was reset to zero. The absorbance of the sample was measured and recorded. The concentration of the reducing sugar can be known by taking the measured absorption value into the previously prepared calibration line.

During the fermentation process, the products (ethanol, acetic acid) metabolized by the cells were quantitatively analyzed by gas chromatography. The procedure is as follows: the fermentation broth is centrifuged at 13300 rpm for 5 minutes, the supernatant is taken, filtered through a 0.2 μm needle filter, and the sample is injected into the gas chromatograph through a auto-injector (Hewlett Packard 7673) (Hewlett) Packard HP 5890 Series II) was used to measure the concentration of the fermentation product. The capillary column in the gas chromatograph is DP-FFAP (30m×0.32mm×0.25 μm ), the initial temperature is 50°C, and the temperature is maintained for 4 minutes, then the temperature is raised to 100°C at 20°C/min for 1 minute. Then, the temperature was raised to 170 ° C at 30 ° C / min for 2.5 minutes; the injection hole and flame ion detector (FID) temperature were both 225 ° C, the split ratio was 10; the carrier gas was nitrogen; The amount of the injected sample was 1 μL.

Among them, regarding the determination of carbon dioxide, the gas to be tested is sucked out from the fermented anaerobic bottle by a syringe, and is put into a 1.8 ml GC sample bottle for concentration, and then the sample is taken from the GC bottle by the GC gas phase needle. 1 ml was taken out and measured in a carbon dioxide sensor (SENTRY ST303).

In addition, regarding the pH measurement, the pH meter is first calibrated. After the calibration is completed, a small amount of the bacterial liquid is taken out to the microcentrifuge tube, and the pH is measured in the pH liquid.

Example 6

For recombinant plastid construction of transketolase and transaldolase, the recombinant plastid construct (pTA) of transketolase is pCDFDuet-1, which is amplified by PCR. The gene fragment of transketolase (TktA) on the chromosome of BL21(DE 3) was cloned to pCDFDuet-1 plastid to form recombinant plastid pTA; and transaldolase (TalB) was recombined in the same manner. Plastid construction (pTB).

In the experiment, in order to allow JB/pTA and JB/pTB with three plastids to be expressed, IPTG (0.5 mM) was selectively added at the second hour of culture to induce a T7 promoter, and the amount of protein was increased. many. During the culture, the experiment was carried out under the conditions of LB as a medium, glucose sugar as a carbon source, 100 mM mops as a pH buffer, and complete anaerobic culture under the conditions of 37 ° C and a rotation speed of 200 rpm. Table 1 shows the strains and plastids used in the experiment.

Table 1. Species and plastids used in the experiment

Results and discussion

Please refer to the first figure. The first figure shows the growth curves of various strains only under aerobic conditions in LB medium. As can be seen from the first figure, the OD ₆₀₀ values of the strains were not significantly different within 10 hours from the beginning, and after 30 hours, the diploid Escherichia coli (JA, JB) showed a higher OD ₆₀₀ value than the wild type. (WT) E. coli was 17.1% lower. In contrast, please refer to the second figure. The second figure shows the (A) growth curve and (B) sugar content of various strains in the aerobic condition of LB medium containing arabinose.

Comparing the first figure with the second (A) figure, it can be found that the OD ₆₀₀ value of each strain showed little difference in the first 10 hours, and the OD ₆₀₀ value was obvious between 10 and 30 hours. Significantly increased (especially J2, J3, JA and JB strains), and further, after 30 hours, or even 100 hours, it can be found that the OD ₆₀₀ value of J1 is similar to that of WT, and the other strains ( The OD ₆₀₀ values of J2, J3, JA and JB) are much larger than WT. It can be clearly seen from the second figure that the dry weight of cells after fermentation of J2, J3, JA and JB strains is 200%-300% higher than that of WT; It can be clearly inferred that the OD ₆₀₀ value of the E. coli strain with the mutated ribulose bisphosphate carboxylase/oxygenase (rbcLS) is significantly better, indicating that the ribulose bisphosphate is mutated. The enzyme/oxygenase (rbcLS) can be grown into E. coli to make it more efficient, and can significantly increase cell dry weight and carbon source metabolism.

At the same time, please also refer to the second (B) chart. The initial arabinose content is 20g/L. In 10 hours, the sugar consumption of each strain is similar; however, between 10 and 30 hours, sugar consumption Significantly increased the amount (especially J2, J3, JA and JB strains), after 30 hours, or even 100 hours, it can be found that the sugar consumption of J1 is similar to that of WT, and the other strains (J2) The sugar consumption of J3, JA and JB) is significantly greater than that of WT; it can be clearly inferred that the Escherichia coli strain with the mutated ribulose bisphosphate carboxylase/oxygenase (rbcLS) can increase the fermentation rate, The fermentation efficiency is increased and the sugar consumption is significantly increased.

Please refer to the third figure. The third picture shows the carbon dioxide produced by fermentation of Escherichia coli. Schematic diagram of the ratio with acetic acid, carbon dioxide and ethanol. As shown, the ratio of carbon dioxide to acetic acid in wild-type E. coli is much higher than that of other plants, indicating that the carbon dioxide of wild-type plants does not have the ability to fix carbon dioxide, thus producing a lot of carbon dioxide. At the same time, the ratio of carbon dioxide to ethanol in wild plants is also higher than that of JB, J1 and J2. It can be seen that the large amount of mutant enzymes in E. coli helps to fix carbon dioxide and reduce carbon dioxide emissions.

The fourth graph is a columnar comparison of carbon dioxide per unit sugar consumption in each strain of the fermentation tank. It can be seen from this figure that both JB/pTA and JB/pTB or JB/pTA+IPTG and JB/pTB+IPTG are less than E.coli BL21(DE3) and JB, and all strains are used when sugar consumption is completed. Glucose has been depleted, thus demonstrating that JB/pTA, JB/pTB, JB/pTA+IPTG and JB/pTB+IPTG are indeed much lower than JB's CO2 emissions per unit of sugar consumption, also demonstrating the use of tktA. The performance of the gene and talB gene can indeed reduce the carbon dioxide emitted by JB.

The fifth graph is a bar graph comparing the yield of ethanol and acetic acid per unit sugar consumption in each strain of the fermentation tank. As can be seen from the figure, JB/pTA, JB/pTB, JB/pTA+IPTG, and JB/pTB+IPTG are less than E.coli BL21(DE3), and are equal to or less than JB, thus indicating JB and JB/pTA, JB. Compared with E.coli BL21 (DE3), /pTB, JB/pTA+IPTG and JB/pTB+IPTG can effectively return the emitted carbon dioxide to the original system and use it, and also make a part of the carbon. Further production of ethanol or acetic acid achieves an effect of increasing the amount of the product.

The sixth graph is a bar graph comparing the CO2 emissions per unit of ethanol and acetic acid production in each strain of the fermentation tank. From the aspects of carbon dioxide emissions/ethanol + acetic acid production, JB/pTA and JB/pTB are 6.25% and 15.6% less than E.coli BL21(DE3) and JB, so it can be explained that every time 1 mole of product is produced, JB/pTA and JB/pTB, The emission of carbon dioxide is less than that of E.coli BL21 (DE3) and JB. It also shows that both JB/pTA and JB/pTB can recycle the carbon dioxide emitted by themselves, which reduces the by-product (carbon dioxide) of the whole system.

In summary, the microorganism of the present invention for forming a fermentation product by fermentation is provided with the following advantages:

(1) Microorganisms containing a mutated enzyme sequence can increase the production efficiency of the fermentation product, and fix carbon dioxide to reduce carbon dioxide emissions.

(2) Increasing the proportion of hydrogen in the gas phase product can reduce the cost of hydrogen purification.

(3) A large number of recombinant protein enzymes can significantly increase cell dry weight and carbon source metabolism in an aerobic or anaerobic environment.

(4) The plastids constructed by the present invention all have the function of reusing carbon dioxide; in other words, the most products can be produced by fermentation with the least amount of carbon dioxide emissions.

Other implementation

All features disclosed in this disclosure can be combined in any manner. Features disclosed in this specification can be replaced with features of the same, equivalent or similar purpose. Therefore, the features disclosed in this specification are one of a series of equivalent or similar features.

In addition, according to the disclosure of the present specification, those skilled in the art can easily make appropriate changes and modifications to different methods and situations without departing from the spirit and scope of the present invention. The implementation aspect is also included in the scope of the patent application.

Claims (9)

A microorganism for fermenting and immobilizing carbon dioxide, comprising a plastid or chromosome comprising a mutated or non-mutated enzyme sequence, the mutated or non-mutated enzyme comprising a plastid selected from the following sequence structure Or a group consisting of: pACYC184-PrkA/pET-rbcLSm1/tktA (JB/PTA) and pACYC184-PrkA/pET-rbcLSm1/talB (JB/pTB); wherein the receptor is a six-carbon sugar, one A hexasaccharide derivative or a combination thereof; wherein the carbon dioxide emission per unit C2 organic matter consumption after fermentation of the plastid or chromosome having the sequence structure is reduced by 6.25%-15.6% compared with the wild type strain. The microorganism of claim 1, wherein the microorganism is a fungus. The microorganism according to claim 2, wherein the fungus is a genus of yeast. The microorganism of claim 1, wherein the microorganism is a bacterium or an archaea or a combination thereof. The microorganism according to claim 4, wherein the bacterium is Escherichia coli, Clostridium acetobutylicum, Bacillus subtilis, Pseudomonas Putia), cyanobacteria, Cupriavidus spp. or Zymomonas mobilis or a combination thereof. The microorganism of claim 1, wherein the microorganism is an alga. The microorganism according to claim 1, wherein the hexose sugar derivative is a hexasaccharide-polymerized disaccharide, oligosaccharide or polysaccharide. The microorganism according to claim 1, wherein the microorganism is subjected to fermentation engineering in a half anaerobic environment. The microorganism according to claim 1, wherein the microorganism is subjected to a fermentation process in a liquid fermentation tank.