CN112877264B - Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid - Google Patents
Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid Download PDFInfo
- Publication number
- CN112877264B CN112877264B CN202110398624.3A CN202110398624A CN112877264B CN 112877264 B CN112877264 B CN 112877264B CN 202110398624 A CN202110398624 A CN 202110398624A CN 112877264 B CN112877264 B CN 112877264B
- Authority
- CN
- China
- Prior art keywords
- pseudomonas putida
- mutant strain
- nicotinic acid
- application
- acid
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
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
- C12P17/00—Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms
- C12P17/10—Nitrogen as only ring hetero atom
- C12P17/12—Nitrogen as only ring hetero atom containing a six-membered hetero ring
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid. The pseudomonas putida mutant strain is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation date: 2021, 3/8, storage address: no. 3 of Beijing West Lu No.1 of Chaoyang district, the preservation number is CGMCC No. 21876. The mutant strain Pseudomonas putida S14 mut-5 of the invention has the activity of 2.68 U.mL‑1It is 2.4 times of wild type. The strain has obviously improved substrate tolerance, can be combined with nitrilase for application, and can convert 3-cyanopyridine for accumulation to obtain 54.5 g.L of 6-hydroxynicotinic acid‑1The optimum pH value is 6.0, and compared with most of niacin dehydrogenases which prefer neutrality and alkalinity, the method is more suitable for industrial production conditions and has wide development prospect in industrial application.
Description
Technical Field
The invention relates to a pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid, belonging to the technical field of microbial fermentation.
Background
In recent years, 6-hydroxynicotinic acid has been receiving much attention as a high-value intermediate for agriculture and chemical industry. It can synthesize low-toxic pyridine methylamine kind of agricultural chemicals and inhibit plant fungus diseases effectively. In the field of materials, 6-hydroxynicotinic acid can participate in the reaction as a unique material, and the derivative thereof can be used as a bridging unit to complete energy transfer. However, 6-hydroxynicotinic acid is difficult to synthesize by chemical methods rapidly and efficiently. At present, the 6-hydroxynicotinic acid is usually produced by taking fumaric acid or malic acid as a raw material through a multi-step synthesis method, wherein the high temperature and strong acid and strong base which are needed can cause environmental pollution and increase the cost, and meanwhile, the product yield is not high, so that the wide application of the product is limited. Therefore, the biotransformation of nicotinic acid by nicotinic acid dehydrogenase to synthesize 6-hydroxynicotinic acid has great potential and commercial value and is receiving great attention.
Niacin dehydrogenase can act to varying degrees on niacin and various niacin analogs. The essence of the hydroxylation process of nicotinic acid is dehydrogenation rather than oxidation, since the O atom in the-OH group is derived from H2O instead of oxygen in air. Niacin dehydrogenases from different strains have different metabolic pathways for converting niacin, but 6-hydroxyniacin is always the first product of all pathways. It has been reported that most nicotinic acid dehydrogenases have an optimum temperature of about 30 to 50 ℃, an optimum pH of about 7.0, and a substrate tolerance of about 10 g.L-1Examples of the nicotinic acid dehydrogenase include Pseudomonas fluorescens TN5 and Pseudomonas putida H9. A high activity (0.58U. mL) was found in 2006 from Pseudomonas putida NA-1-1) The nicotinic acid dehydrogenase of (1). In 2007, a nicotinic acid dehydrogenase is also discovered in pseudomonas BK-1, and the enzyme activity of the nicotinic acid dehydrogenase is 0.57 U.mL-1. In 2008, it was found that the nicotinic acid dehydrogenase activity of Pseudomonas putida BKC4 was 0.53U mL-1. In 2009, Pseudomonas putida KT2440 nicotinic acid dehydrogenase activity was reported to be 0.34 U.mL-1. In 2017, Pseudomonas putida H9 separated from rotten fish can convert nicotinic acid into 6-hydroxynicotinic acid with activity of 0.37 U.mL-1. It was reported that P.fluoroscens TN5 could convert 1.4M nicotinic acid to 191 g.L within 45h-16-Hydroxynicotinic acid, space-time yield 0.57g (g.h)-1。
However, the performance of wild bacteria generally cannot meet the industrial requirements, so that various strategies are generally required to improve the catalytic performance of the biocatalyst.
Disclosure of Invention
In order to solve the technical problems, the invention excavates a high-yield nicotinic acid dehydrogenase P.putida S14 from soil, applies the high-yield nicotinic acid dehydrogenase P.putida S14 to the production of 6-hydroxynicotinic acid, and identifies the product. Performing ARTP mutagenesis on P.putida S14 wild bacteria, performing high-throughput screening by adopting a high-throughput liquid transfer workstation and a high-throughput microorganism cloning screening system to obtain a mutant strain with higher enzyme activity, performing detection on the most suitable pH, substrate tolerance and passage stability, and performing cascade application with nitrilase to produce 6-hydroxynicotinic acid.
The first purpose of the invention is to provide a pseudomonas putida mutant strain, which is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms with the preservation date: 2021, 3/8, storage address: no. 3 of Beijing West Lu No.1 of Chaoyang district, the preservation number is CGMCC No. 21876.
The second purpose of the invention is to provide the application of the pseudomonas putida mutant strain in catalyzing nicotinic acid to produce 6-hydroxy nicotinic acid.
Further, the application specifically comprises the step of producing the nicotinic acid dehydrogenase by adopting the pseudomonas putida mutant strain through fermentation, and the step of catalyzing nicotinic acid to produce 6-hydroxynicotinic acid by using pseudomonas putida mutant strain cells or the nicotinic acid dehydrogenase as a catalyst.
Furthermore, in the application, the concentration of a substrate nicotinic acid is 30-40 g.L-1。
Furthermore, the catalysis condition is that the temperature is 28-35 ℃ and the pH value is 5.5-7.0.
The third purpose of the invention is to provide the application of the pseudomonas putida mutant strain in catalyzing 3-cyanopyridine to produce 6-hydroxynicotinic acid, wherein the application is that the nicotinic acid dehydrogenase produced by fermentation of the pseudomonas putida mutant strain and nitrilase perform enzymatic cascade reaction, and 3-cyanopyridine is used as a substrate to produce 6-hydroxynicotinic acid.
Further, the nitrilase is derived from pseudomonas putida with the preservation number of CGMCC No. 14276.
Further, the application specifically comprises that the pseudomonas putida mutant strain and the pseudomonas putida with the preservation number of CGMCC No.14276 are used as catalysts, and20～30g·L-13-cyanopyridine is used as a substrate to catalyze and produce 6-hydroxynicotinic acid.
Furthermore, the catalysis condition is that the temperature is 26-32 ℃ and the pH value is 6.0-7.0.
Furthermore, the addition amount of the pseudomonas putida mutant strain and the pseudomonas putida with the preservation number of CGMCC No.14276 is 1: 0.5-2 in weight ratio.
The invention has the beneficial effects that:
the invention obtains a high-yield nicotinic acid dehydrogenase strain Pseudomonas putida S14 through screening, and the activity of the strain is 1.11 U.mL-1The optimum pH is 5.5, and the yield of 6-hydroxynicotinic acid is 176 g.L-1The space-time yield is 1.36g (g.h)-1. Through ARTP mutagenesis and obtaining a mutant strain Pseudomonas putida S14 mut-5 with remarkably improved enzyme activity, the activity is 2.68 U.mL-1It is 2.4 times of wild type. The strain has obviously improved substrate tolerance, can be combined with nitrilase for application, and can convert 3-cyanopyridine for accumulation to obtain 54.5 g.L of 6-hydroxynicotinic acid-1The optimum pH value is 6.0, and compared with most of niacin dehydrogenases which prefer neutrality and alkalinity, the method is more suitable for industrial production conditions and has wide development prospect in industrial application.
Microbiological material preservation information: pseudomonas putida S14 mut-5, taxonomic name Pseudomonas putida, deposited in China general microbiological culture Collection center, with the deposition date: 2021, 3/8, storage address: no. 3 of Xilu No.1 of Beijing, Chaoyang, China academy of sciences, microbiological research institute, preservation number CGMCC No. 21876.
Description of the drawings:
FIG. 1 shows the screening of nicotinic acid dehydrogenase activity;
FIG. 2 shows the fed batch synthesis of 6-hydroxynicotinic acid from Pseudomonas putida S14 using nicotinic acid as substrate;
FIG. 3 shows LC-MS analysis of the conversion product; a is a 6-hydroxynicotinic acid standard product, and b is a conversion product;
FIG. 4 is a graph showing the lethality of Pseudomonas putida S14 subjected to ARTP mutagenesis;
FIG. 5 shows a positive mutation selection of Pseudomonas putida S14 mutant;
FIG. 6 is the optimum pH of Pseudomonas putida S14 and mut-5;
FIG. 7 shows the substrate tolerance of Pseudomonas putida S14 and mut-5;
FIG. 8 is the genetic stability of mut-5;
FIG. 9 shows the batch feeding synthesis of 6-hydroxynicotinic acid by mut-5 using nicotinic acid as a substrate;
FIG. 10 is a schematic representation of mut-5 sequential conversion of 3-cyanopyridine to 6-hydroxynicotinic acid;
FIG. 11 shows the synthesis of 6-hydroxynicotinic acid by successive conversion of 3-cyanopyridine with Pseudomonas putida S14.
Detailed Description
The present invention is further described below in conjunction with specific examples to enable those skilled in the art to better understand the present invention and to practice it, but the examples are not intended to limit the present invention.
Example 1:
this example illustrates a method for screening a bacterium producing nicotinic acid dehydrogenase.
1mL of the supernatant of the soil sample was added to 100mL of the enrichment medium and reacted at 30 ℃ and 200rpm for 20 hours. After enrichment of the microorganisms, 1mL of the culture was transferred to selection medium and incubated for 72h at 30 ℃. The culture was then diluted 10-6-10-8After doubling, the cells were plated on separate medium plates and incubated at 30 ℃ for 72 hours to isolate single colonies.
Single colonies were inoculated into 96-well plates containing 100. mu.L of fermentation medium and cultured at 30 ℃ and 200rpm for 20 h. Then, using the property that 6-hydroxynicotinic acid can be developed under ultraviolet light, 3. mu.L of fermentation supernatant was spotted on filter paper, respectively, and irradiated under ultraviolet light of 300nm to perform primary screening. Selection medium was transferred to 24-well plates containing fresh fermentation medium and incubated at 30 ℃ and 200rpm for 24 h. The activity of the nicotinic acid dehydrogenase is determined by an enzyme activity detection method based on an enzyme-labeling instrument. Through the combination of development screening and enzyme activity determination, a strain marked as S14 is obtained, and the enzyme activity of the strain can reach 1.11 U.mL-1(FIG. 1).
The obtained wild strain is identified, the 16S rDNA sequence of the wild strain is amplified, the length is 1403bp, and then the wild strain is compared with a report sequence in an NCBI public database. The result shows that the strain has higher sequence homology with pseudomonas putida (AY395005.1, similarity is 99.93%). Therefore, this strain was named Pseudomonas putida S14.
Example 2:
this example illustrates the batch fed synthesis of 6-hydroxynicotinic acid by Pseudomonas putida S14 using nicotinic acid as substrate.
The 6-hydroxynicotinic acid is synthesized by biotransformation of nicotinic acid with the resting cell of pseudomonas putida S14 which produces nicotinic acid dehydrogenase as catalyst. Nicotinic acid was added every 2h at 30 ℃ to maintain the substrate concentration at 30 g.L-1Then, the concentration of 6-hydroxynicotinic acid accumulated by 14 substrate feeds in 30 hours reaches 176 g.L-1(FIG. 2), wherein the dry weight of resting cells was 4.3 g.L-1. The space-time yield of Pseudomonas putida S14 nicotinic acid dehydrogenase was 1.36g (g.h)-1The method is the highest level of the production efficiency of the 6-hydroxynicotinic acid reported in the literature at present.
The molecular weight of the converted product was determined by LC-MS to be 139, and the peak-off time was consistent with that of the 6-hydroxynicotinic acid standard (FIG. 3).
Example 3:
this example illustrates the method of breeding high-yield nicotinic acid dehydrogenase by ARTP mutagenesis.
Pseudomonas putida S14 was cultured to log phase (OD)6000.6-0.8), 15 μ L of the pretreated cell suspension was spread evenly on a metal slide and irradiated under plasma beam for 0, 5, 10, 15, 25, 45, 65s, respectively. Slides were transferred to EP tubes containing 1mL NaCl (154mM, pH 7.0) and shaken immediately for 2 min. Finally, the mutagenic solution of the mutant strain was diluted and spread and cultured at 30 ℃ for 20-24 h. Drawing a lethality curve, and calculating a formula:
Z＝(1-X/X0)×100％
z is lethality, X is the number of mutant colonies, X0Is the number of control colonies.
90% lethality was set as the mutation standard. When the treatment time was 10s, the lethality reached 92.16% (FIG. 4). When the treatment time exceeded 65s, all cells lost activity. Thus, ARTP mutagenesis was performed for 10 seconds.
Example 4:
this example illustrates a high throughput screening method for positive mutations in nicotinic acid dehydrogenase.
A high throughput screening method was established using the high throughput microbial clonal screening system (Molecular Devices Qpix 420). The single colony of the target was picked into a 48-well plate using an automatic bacteria picker and cultured at 30 ℃ for 21 hours. The well plate was centrifuged to collect resting cells, and after the resting cells were washed 2 times by pipetting through the arm, 1mL of 10 g.L was added-1Nicotinic acid. Then, the conversion reaction was carried out at 30 ℃ for 1 hour, and the nicotinic acid dehydrogenase activity of the mutant was examined. By the method, 7680 mutants are screened together, 9 mutants (with repeated strain removed) with activity improved by 150-240% compared with the wild type are selected for rescreening, wherein the mutant strain mut-5 has the highest catalytic activity of 2.68 U.mL-12.4 times of wild type (figure 5), and is currently preserved in China general microbiological culture Collection center with the preservation number of CGMCC No. 21876.
The enzyme activity detection method comprises the following steps: the fermentation broth was centrifuged at 12000rpm for 2min and the resting cells were collected after washing 2 times with PBS buffer (20mM, pH 7.0). 1mL of 10 g.L was added-1Nicotinic acid solution, at 30 ℃, 200rpm for 1 h. After centrifugation at 12000rpm for 2min, the absorbance of the reaction mixture supernatant at 295nm was measured using a microplate reader (Multiskan assay, Molecular Devices, USA).
One unit (U) of niacin dehydrogenase activity is defined as: the amount of enzyme required to produce 1. mu. mol of 6-hydroxynicotinic acid in 1min [7 ]. The calculation formula is as follows:
1U＝n×m×106/M×t
wherein n is the dilution factor, and m is the mass concentration (g.L) of 6-hydroxynicotinic acid-1) M is the molar mass of 6-hydroxynicotinic acid 139.11, and t is the reaction time (min).
Example 5:
this example illustrates the method of detection of optimal pH and substrate tolerance.
The pH of the mixture was adjusted to 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, and 9.0 at a concentration of 10 g.L-1Taking 1mL of resting cells, reacting at 30 ℃ for 1h, measuring the enzyme activity according to the medium enzyme activity detection method in the embodiment 2, and drawing an optimal pH curve. As shown in FIG. 3, the optimal pH for mut-5 is 6.0 (FIG. 6).
The pH of the mixture was 7.0, and the concentrations were 5, 10, 20, 30, 40, 50 g.L-1The enzyme activity of the nicotinic acid solution was measured as described above according to the method for detecting medium enzyme activity in example 2, and a substrate tolerance curve was plotted. As shown in FIG. 7, the substrate tolerance of the wild type was 30 g.L-1While the substrate tolerance of mut-5 is obviously improved to 40 g.L-1。
Example 6:
the examples illustrate the method of testing the stability of the passage.
Inoculating the mutant strain with high enzyme activity obtained by screening into a fermentation culture medium, and culturing for 20h at 30 ℃. And (3) taking 1mL of bacterial liquid to measure the enzyme activity, and carrying out passage for 15 times in a fermentation medium to examine the passage stability. As can be seen from FIG. 8, mut-D3 has good passaging stability.
Example 7:
this example illustrates the batch fed synthesis of 6-hydroxynicotinic acid by Pseudomonas putida S14 mut-5 using nicotinic acid as substrate.
The 6-hydroxynicotinic acid is synthesized by biotransformation of nicotinic acid with Pseudomonas putida S14 mut-5 resting cells producing nicotinic acid dehydrogenase as catalyst. As shown in FIG. 9, nicotinic acid was fed every 2 hours at 30 ℃ to maintain the substrate concentration at 40 g.L-1The concentration of 6-hydroxynicotinic acid accumulated in 38 hours is 244.9 g.L-1This is 139% of the wild type transformation yield.
Example 8:
this example illustrates the establishment of a nitrilase-nicotinic acid dehydrogenase cascade.
A nitrilase-nicotinic acid dehydrogenase cascade system is established, and 3-cyanopyridine is used as a substrate to produce 6-hydroxynicotinic acid.
Pseudomonas putida mut-D3 (preservation number is CGMCC No.14276) is inoculated in LB culture medium and cultured for 36h at 30 ℃. P. putida S14 inoculated in fermentation MediumCulturing at 30 deg.C for 21 hr, and culturing in culture medium (g.L)-1) Comprises the following steps: tryptone 10, nicotinic acid 10, yeast extract 5, MgSO4·7H2O 5，MnSO4·7H2O 5，ZnSO4·7H2O 5，CaCl2·H2O5, molybdic acid 2, K2HPO4 1，CoCl2·6H2O0.1 obtaining Pseudomonas putida mut-D3 and P.putida S14 mut-5 resting cells respectively.
Nitrilase-producing pseudomonas putida mut-D3 and p.putida S14 mut-5 were used as catalysts, resting cells were collected and cultured in a 1: 1 was resuspended in 30 g.L-1In the 3-cyanopyridine solution at 30 ℃ and as shown in FIG. 10, 6-hydroxynicotinic acid accumulated to 54.5 g.L in 585min after 15 batches of feeding-1。
For comparison, nitrilase-producing pseudomonas putida mut-D3 and p.putida S14 were used as catalysts, resting cells were collected and cultured in a 1: 1 was resuspended in 30 g.L-1At 30 ℃ as shown in FIG. 11, 6-hydroxynicotinic acid was accumulated to 31.2 g.L in 330min-1This is probably because the substrate 3-cyanopyridine exerts a deleterious effect on cells, reducing the stability of p.
The above-mentioned embodiments are merely preferred embodiments for fully illustrating the present invention, and the scope of the present invention is not limited thereto. The equivalent substitution or change made by the technical personnel in the technical field on the basis of the invention is all within the protection scope of the invention. The protection scope of the invention is subject to the claims.
Claims (10)
1. A pseudomonas putida mutant strain is characterized by being named as pseudomonas putida (A)Pseudomonas putida) The culture medium is preserved in China general microbiological culture Collection center, and the preservation date is as follows: 2021, 3/8, storage address: beijing, Chaoyang district, Beichen Xilu No.1 Hospital No. 3 with preservation number of CGMCC No. 21876.
2. The use of the mutant strain of pseudomonas putida of claim 1 for catalyzing the production of 6-hydroxynicotinic acid from nicotinic acid.
3. The application of claim 2, wherein the application is specifically to produce the nicotinic acid dehydrogenase by using the pseudomonas putida mutant strain for fermentation, and the 6-hydroxynicotinic acid is produced by catalyzing nicotinic acid with the pseudomonas putida mutant strain cells or the nicotinic acid dehydrogenase as a catalyst.
4. The use of claim 2, wherein the concentration of nicotinic acid as a substrate is 30-40 g.L-1 。
5. The use according to claim 4, wherein the catalysis conditions are a temperature of 28-35 ℃ and a pH of 5.5-7.0.
6. The use of the mutant pseudomonas putida strain of claim 1 for catalyzing 3-cyanopyridine to produce 6-hydroxynicotinic acid, wherein the use is to perform an enzymatic cascade reaction of a nicotinic acid dehydrogenase produced by fermentation of the mutant pseudomonas putida strain and a nitrilase, and produce 6-hydroxynicotinic acid by using 3-cyanopyridine as a substrate.
7. The use according to claim 6, wherein said nitrilase is derived from Pseudomonas putida having the accession number CGMCC number 14276.
8. The application of claim 6, wherein the application specifically comprises the pseudomonas putida mutant strain and the pseudomonas putida with the preservation number of CGMCC number 14276 as catalysts, and 20-30 g-L-13-cyanopyridine is used as a substrate to catalyze and produce 6-hydroxynicotinic acid.
9. The use according to claim 8, wherein the catalysis conditions are a temperature of 26-32 ℃ and a pH of 6.0-7.0.
10. The use of claim 8, wherein the weight ratio of the pseudomonas putida mutant strain to the pseudomonas putida with the preservation number of CGMCC number 14276 is 1: 0.5-2.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110398624.3A CN112877264B (en) | 2021-04-12 | 2021-04-12 | Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110398624.3A CN112877264B (en) | 2021-04-12 | 2021-04-12 | Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112877264A CN112877264A (en) | 2021-06-01 |
| CN112877264B true CN112877264B (en) | 2022-04-05 |
Family
ID=76040074
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110398624.3A Active CN112877264B (en) | 2021-04-12 | 2021-04-12 | Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112877264B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117070417A (en) * | 2023-09-07 | 2023-11-17 | 河南大学 | Pseudomonas grass HD530 for producing 6-hydroxynicotinic acid and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0253438A2 (en) * | 1986-07-08 | 1988-01-20 | Shell Internationale Researchmaatschappij B.V. | New strains of Pseudomonas putida and their use |
| CN102337233A (en) * | 2011-05-06 | 2012-02-01 | 江南大学 | Pseudomonas putida and method for producing nicotinic acid or isonicotinic acid through converting Pseudomonas putida |
| CN112011490A (en) * | 2020-09-16 | 2020-12-01 | 安徽瑞邦生物科技有限公司 | Pseudomonas putida strain and application thereof in degrading nicotinic acid |
-
2021
- 2021-04-12 CN CN202110398624.3A patent/CN112877264B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0253438A2 (en) * | 1986-07-08 | 1988-01-20 | Shell Internationale Researchmaatschappij B.V. | New strains of Pseudomonas putida and their use |
| CN102337233A (en) * | 2011-05-06 | 2012-02-01 | 江南大学 | Pseudomonas putida and method for producing nicotinic acid or isonicotinic acid through converting Pseudomonas putida |
| CN112011490A (en) * | 2020-09-16 | 2020-12-01 | 安徽瑞邦生物科技有限公司 | Pseudomonas putida strain and application thereof in degrading nicotinic acid |
Non-Patent Citations (2)
| Title |
|---|
| Desta M ; Liu G ; Hu H.Purification, crystallization and preliminary X-ray analysis of NicX, a key enzyme in nicotinic acid degradation from Pseudomonas putida KT2440.《Journal of biological regulators and homeostatic agents》.2019, * |
| 恶臭假单胞菌 NA-1菌体培养转化和静息细胞转化联合工艺生产6-羟基烟酸研究;徐莉 等;《微生物学报》;20060204;第63-67页，参见全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112877264A (en) | 2021-06-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US8900836B2 (en) | Acid production by fermentation | |
| CN109628339B (en) | L-lactic acid producing strain and method for producing L-lactic acid by using same | |
| US10196660B2 (en) | Xylene monooxygenase-producing strain Arthrobacter woluwensis and its application | |
| JP2017502677A (en) | Sporolactobacillus terae and its use | |
| CN112239738B (en) | Escherichia coli capable of producing succinic acid and application thereof | |
| CN112877264B (en) | Pseudomonas putida mutant strain and application thereof in biological preparation of 6-hydroxynicotinic acid | |
| Jiang et al. | Bioproduction of hydrogen by simultaneous saccharification and fermentation of cassava starch with 2-deoxyglucose-resistant mutant strains of Clostridium tyrobutyricum | |
| CN112680484B (en) | Method for producing 3, 4-dihydroxybutyric acid by using double-bacterium co-culture system | |
| CN108384730B (en) | Lactobacillus paracasei and application thereof in conversion synthesis of phenyllactic acid | |
| CN114606142B (en) | L-malic acid producing strain with high production efficiency, method and application | |
| KR20050022591A (en) | Method for Producing Bacterial Cellulose Using the Waste of Beer Fermentation Broth | |
| Ryu et al. | Characteristics and glycerol metabolism of fumarate‐reducing Enterococcus faecalis RKY1 | |
| KR101402659B1 (en) | Actinobacillus succinogenes AND MANUFACTURING METHOD OF SUCCINIC ACID USING THE SAME | |
| Hussain et al. | Enhancement effect of AgO nanoparticles on fermentative cellulase activity from thermophilic Bacillus subtilis Ag-PQ | |
| CN110982757A (en) | Enterobacter cloacae ZJPH1903 and application | |
| Ikram et al. | Evaluation, Characterization and Molecular Analysis of Cellulolytic Bacteria from Soil in Peshawar, Pakistan | |
| KR20200038377A (en) | An Escherichia coli adapted to methanogenic co-culture and producing succinate from waste glycerol by using adapted Escherichia coli | |
| CN111500486B (en) | Strain capable of directly synthesizing butanol by using inulin as unique carbon source and application thereof | |
| CN113215064B (en) | Slime bacterium for producing meishadazole compounds and application thereof | |
| KR100355496B1 (en) | A novel obligately symbiotic thermophile Symbiobacterium toebii SC-1 producing thermostable L-tyrosine phenol-lyase and L-tryptophan indole-lyase and a method for screening its relative bacteria | |
| CN114606278B (en) | Method for synthesizing 2,5-furandicarboxylic acid by using methyl bacillus catalysis | |
| CN117070417A (en) | Pseudomonas grass HD530 for producing 6-hydroxynicotinic acid and application thereof | |
| US8771996B2 (en) | Marine bacterium of metabolizing 3,6-anhydro-L-galactose and use of the same | |
| Cha | Biological production of optically active muconolactones by Rhodococcus rhodochrous | |
| HOMMA et al. | Preparation of high-optical-purity D-lactic acid from DL-lactic acid using optical isomer preferential lactic acid assimilating microorganisms |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |