CN101760485A - Preparation method of novel biomaterial polyhydroxyl alkanoic acid - Google Patents
Preparation method of novel biomaterial polyhydroxyl alkanoic acid Download PDFInfo
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- CN101760485A CN101760485A CN200810162764A CN200810162764A CN101760485A CN 101760485 A CN101760485 A CN 101760485A CN 200810162764 A CN200810162764 A CN 200810162764A CN 200810162764 A CN200810162764 A CN 200810162764A CN 101760485 A CN101760485 A CN 101760485A
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Abstract
The invention discloses a preparation method of novel biomaterial polyhydroxyl alkanoic acid. Al-caligenes eutrophus is adopted as original strain, and organic acid with various micromolecules is formed by utilizing organic waste through anaerobic acidification, and the organic acid is used as a carbon source for producing PHAs (polyhydroxyalkanoicacids), thereby not only the production cost is greatly lowered, but also the higher cell dry mass and PHAs yield of the strain can be obtained through a mode of feeding fermentation. Meanwhile, the method provides a good direction for the processing of the organic waste, thereby not only the problem of processing the organic waste is solved, but also the environment is optimized, and the produced product polyhydroxyl alkanoic acid has good and wide application prospects.
Description
Technical field
The invention relates to a kind of preparation method of novel biomaterial polyhydroxyl alkanoic acid, belong to field of novel or bioengineering field.
Background technology
Polyhydroxyl alkanoic acid (PHAs) is that a class is by biosynthetic high molecular polymer, physicochemical properties such as it has not only that light specific gravity, Stability Analysis of Structures, plasticity-are strong, optical activity and piezoelectricity are good, and have biological characteristicses such as biodegradability, biological recyclability, Bc, degraded product nontoxicity, be the environmentally friendly novel biomaterial of a class therefore.
As far back as the twenties in 20th century, people have just found polyhydroxyl alkanoic acid in bacterium.20th century posterior lobe, " white pollution " problem that plastics industry hypergrowth, plastics cause appears suddenly out day by day, is seriously threatening the physical environment of people existence and healthy.In addition. the main production raw material of petrochemical industry plastics---oil is non-renewable natural resources, so scientists begins to seek plastic-substitute, and the physico-chemical property of having found polyhydroxyl alkanoic acid in numerous research materials is close with the petrochemical industry plastics, is one of ideal plastics equivalent material.From then on the development research of PHAs has obtained attention.The seventies in 20th century, the research and development of PHAs become one of world's hot subject.Researcher has carried out a large amount of research from aspects such as microorganism resource, physiological action, pathways metabolism and Regulation Mechanism, fermentation manufacturing technique, physico-chemical property, mechanical propertys and uses thereof, obtained breakthrough progress, made part PHAs product drop into industrialized trial production finally.
Production and the application of PHAs at present has been subjected to great challenge, and the scientific worker concentrates on the seed selection of good production bacterial classification and the optimization aspect of zymotechnique to the emphasis of research.Simultaneously, carried out the research of other modes of production,, opened up the new way of producing PHAs by arable farming as making up transgenic plant; The exploitation immobilized enzyme is set up Production by Enzymes PHAs novel process, reduces production costs scheming.In addition, also carried out the applied research of PHAs on high value added product.These researchs have all obtained progress in various degree.
Nineteen twenty-six, the researchist of Pasteur Institut isolated two kinds of particulate component from bacillus megaterium, confirmed that after deliberation wherein a kind of particulate chemical ingredients is poly--p-hydroxybutyric acid (PHB), has proposed the key concept of this base polymer first.Afterwards, people have also found this base polymer successively in other bacteriums, and prove that gradually the synthetic PHAs of bacterium is a commonplace phenomenon, and having found out in procaryotic more than 90 belong to has 300 various bacteria to have the ability of synthetic PHAs.Like this, bacterium just becomes the preferred material that people research and develop PHAs.
Through the investigation of Biological resources, the PHAs that has filtered out multiple wild-type produces bacterial strain, and the main bacteria seed that is used for fermentative production has really to support has a liking for some bacterial strains that alkali bacterium, pseudomonas and first bacteria etc. belong to.As, alcaligenes eutrophus is that the good PHB of a strain produces bacterial strain, and its available carbon source is extensive, and can add up the PHAs of high level in born of the same parents and widely adopted.Among the synthetic PHAs of Ralstonia eutropha institute, when side chain substituents contained a carbon atom, the monomer of polymkeric substance was called butyric ester, abbreviates HB as, and the formed polymkeric substance of HB is PHB; When the side chain substituents in the PHAs partial monosomy contains 2 carbon atoms, monomer is called hydroxyl valerate, abbreviate HV as, the HV component has very important meaning to the performance that improves PHAs. owing to the reason of microorganism cells metabolic mechanism, HV is often with HB and deposit among the PHAs, the multipolymer that forms is (being called for short PHBV), the ratio of HV component high more (ratio that is HV/PHAs is big more) among the PHAs generally, the performance of PHAs is good more. on the other hand, and owing to the raw materials cost and too high its large-scale application that hindered of extraction cost of PHAs production at present.
It is starting strain that the present invention adopts Ralstonia eutropha, utilize organic waste as the carbon source of producing PHAs, can reduce production costs greatly, the main component of organic waste behind anaerobic acidification is the mixing acid that various micromolecular organic acids (as acetate, propionic acid, lactic acid, butyric acid etc.) are formed, bacterial strain adds fermentation through stream, can obtain higher cell and do matter and PHAs output.
Summary of the invention
One of purpose of the present invention has provided a kind of preparation method of novel biomaterial polyhydroxyl alkanoic acid.
Two of purpose of the present invention is that the processing for organic waste provides a kind of new approach.
Technical solution of the present invention is as follows:
Well-grown Ralstonia eutropha bacterial strain is cultivated in the 500ml triangular flask that the 100ml seed culture medium is housed on the seed culture inoculation one ring slant medium, shakes a bottle rotating speed 200r/min, 30 ℃ of temperature, incubation time 30h.
The bottled liquid 75ml of shake flask fermentation 500ml triangle, inoculum size 10% is shaken a bottle rotating speed 200r/min, and 30 ℃ of temperature are about fermentation time 36h.
Jar fermentation 5L jar mixing speed 400r/min, air flow 4.5L/min, adopt stream to add the mode of fermentation: the canned liquid 2.6L of 5L, inoculum size 10%, stream adds whole process and adds for staged stream stage by stage, and the carbon source concentration of stream liquid feeding is 100g/L, wherein acetate: lactic acid: propionic acid: butyric acid=1: 1: 3: 3, and add nitrogenous source since 12h stream, it is 20g/L that stream adds nitrogen concentration.Regulate the pH value by auto-feeding 3mol/L sodium hydroxide solution and 3mol/L hydrochloric acid soln, about fermentation time 60h.
With chloroform (every 10ml chloroform adds the 0.2g stem cell) broken wall, 60 ℃ of insulation 24h filter the PHAs assay with lyophilize gained cell, and filtrate is dried naturally, is PHAs.
It is starting strain that the present invention adopts Ralstonia eutropha, the main component of organic waste behind anaerobic acidification is the mixing acid that various micromolecular organic acids (as acetate, propionic acid, lactic acid, butyric acid etc.) are formed, utilize organic waste as the carbon source of producing PHAs, not only can reduce production costs greatly, and bacterial strain adds the mode of fermentation through stream, can obtain higher cell and do matter and PHAs output.Meanwhile, this method has not only solved the handling problem of organic waste, and has optimized environment for the processing of organic waste provides a good direction, and the product polyhydroxyl alkanoic acid of producing has good and application prospects.
Claims (3)
1. the preparation method of a novel biomaterial polyhydroxyl alkanoic acid is characterized in that may further comprise the steps:
Well-grown Ralstonia eutropha bacterial strain is cultivated in the 500ml triangular flask that the 100ml seed culture medium is housed on the seed culture inoculation one ring slant medium, shakes a bottle rotating speed 200r/min, 30 ℃ of temperature, incubation time 30h.
The bottled liquid 75ml of shake flask fermentation 500ml triangle, inoculum size 10% is shaken a bottle rotating speed 200r/min, and 30 ℃ of temperature are about fermentation time 36h.
Jar fermentation 5L jar mixing speed 400r/min, air flow 4.5L/min, adopt stream to add the mode of fermentation: the canned liquid 2.6L of 5L, inoculum size 10%, stream adds whole process and adds for staged stream stage by stage, and the carbon source concentration of stream liquid feeding is 100g/L, wherein acetate: lactic acid: propionic acid: butyric acid=1: 1: 3: 3, and add nitrogenous source since 12h stream, it is 20g/L that stream adds nitrogen concentration.Regulate the pH value by auto-feeding 3mol/L sodium hydroxide solution and 3mol/L hydrochloric acid soln, about fermentation time 60h.
With chloroform (every 10ml chloroform adds the 0.2g stem cell) broken wall, 60 ℃ of insulation 24h filter the PHAs assay with lyophilize gained cell, and filtrate is dried naturally, is PHAs.
2. according to said acetate, lactic acid, propionic acid, butyric acid in the right 1, it is characterized in that they come from that organic waste are transformed through anaerobic acidification.
3. according to said PHAs in the right 1, it is characterized in that it is a kind of novel, environmentally friendly biomaterial, is one of ideal plastics equivalent material.
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| CN200810162764A CN101760485A (en) | 2008-12-11 | 2008-12-11 | Preparation method of novel biomaterial polyhydroxyl alkanoic acid |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543297A (en) * | 2016-03-02 | 2016-05-04 | 中国科学院过程工程研究所 | Method for preparing polyhydroxyalkanoate by combining hydrogenogens and alcaligenes eutrophus to convert biomass and CO2 |
| US10358665B2 (en) | 2016-09-08 | 2019-07-23 | Imam Abdulrahman Bin Faisal University | Method for producing polyhydroxyalkanoate by fed-batch culture of bacillus bacteria in a medium containing date syrup |
| US11559604B2 (en) | 2017-08-31 | 2023-01-24 | University Of Westminster | Nerve conduits |
-
2008
- 2008-12-11 CN CN200810162764A patent/CN101760485A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543297A (en) * | 2016-03-02 | 2016-05-04 | 中国科学院过程工程研究所 | Method for preparing polyhydroxyalkanoate by combining hydrogenogens and alcaligenes eutrophus to convert biomass and CO2 |
| US10358665B2 (en) | 2016-09-08 | 2019-07-23 | Imam Abdulrahman Bin Faisal University | Method for producing polyhydroxyalkanoate by fed-batch culture of bacillus bacteria in a medium containing date syrup |
| US11559604B2 (en) | 2017-08-31 | 2023-01-24 | University Of Westminster | Nerve conduits |
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Open date: 20100630 |