CN102453739A - Preparation method of chitosan - Google Patents
Preparation method of chitosan Download PDFInfo
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- CN102453739A CN102453739A CN2010105160348A CN201010516034A CN102453739A CN 102453739 A CN102453739 A CN 102453739A CN 2010105160348 A CN2010105160348 A CN 2010105160348A CN 201010516034 A CN201010516034 A CN 201010516034A CN 102453739 A CN102453739 A CN 102453739A
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Abstract
Belonging to the field of biological fermentation, the invention specifically relates to a preparation method of chitosan. The preparation method of chitosan in the invention has the advantages of simple preparation, short time, and high yield. The method provided in the invention comprises the steps of: aspergillus niger fermentation; chitin preparation.
Description
Technical field:
The invention belongs to the biological fermentation field, in particular, relate to a kind of preparation method of chitosan.
Background technology:
Chitosan is the product of chitin deacetylate after concentrated alkali solution is handled, chemistry (1, the 4)-2-amino-2-deoxidation-β-D-glucose that gathers by name.The biocompatibility of chitosan is good, and is extremely extensive in the application of aspects such as biomedicine and pharmacy, can be used as burn dressing and Wound-healing agent, the wrapping gauze with treatment with chitosan after, speed of wound healing can improve 75%.With the absorbability sutures that chitosan is processed, physical strength is high, but long storage can sterilize with ordinary method, can dye, can mix medicament, can be absorbed by tissue degradation, exempt the misery that the patient takes out stitches.Chitosan can gastric acid inhibitory and ulcer, has the effect of degraded SUV and triglyceride.Heparin is a kind of extremely effectively anti-coagulant that has sulfonic group, carboxyl; The chitosan of sulphating is structurally similar with heparin; This heparitin verivate generally has quite even surpasses heparin activity, for synthetic cheap anti-coagulant is provided valid approach is provided.In addition, chitosan also can be used for making artificial kidney dialysis membrane and contact lens.By the microcapsule that Preparation of Chitosan goes out, be a kind of macromolecule member material of biodegradation type, be good and have the medical slow release system of development prospect.
In addition, chitosan can be used as preservative film on food.Its aqueous solution is applied to fruit and vegetable surfaces, can artificially forms the closed environment of a hypoxemia high carbon dioxide at fruit and vegetable surfaces, suppress fruits and vegetables and breathe, antibacterial breeding simultaneously improves the fruits and vegetables glossiness, improves the organoleptic quality of fruit tree.
The preparation method of chitosan length mostly consuming time, high to equipment requirements, and productive rate is generally very low.
Summary of the invention:
The present invention is exactly to the problems referred to above, and a kind of preparation method who prepares weak point simple, consuming time, chitosan that productive rate is high is provided.
In order to realize above-mentioned purpose of the present invention, preparation process of the present invention is:
1. fermentation of Aspergillus niger
The slant medium preparation: 4% agar, 2% yeast extract paste, 2% peptone, 4% sucrose, the test tube of packing into after the heating and filtering is in 121 ℃ of autoclaving 20min; On the aseptic technique platform, inoculate, cultivate 0～110h in 22～34 ℃ of electro-heating standing-temperature cultivators; Scrape the mould spore of the black marrow of getting on the fresh inclined-plane with transfering loop; Change over to and sterilized water is housed and is equipped with in the tool plug triangular flask of granulated glass sphere; Fully suspension concentration is calculated with blood counting chamber in the vibration back, and is diluted to desired concn (108 spores of 7108～9X/mL) with sterilized water; In the 500mL Erlenmeyer flask, encase the Erlenmeyer flask bottleneck, in 121 ℃ of autoclaving 20min with multilayer gauze and kraft paper; Spore suspension is inserted according to quantity at the aseptic technique platform in the sterilization back, the constant temperature shaker fermentation, and shaking speed is 120～210r/min;
2. the preparation of chitosan
Get fermented liquid, centrifuge washing thalline to supernatant is colourless, and (thalline: alkali lye=1: 10w/V), centrifugal behind 50 ℃ of following processing 3h, it is colourless that water washing is precipitated to supernatant to get gained deposition adding 7% sodium hydroxide solution; Get above-mentioned deposition, add ethanol: (thalline: alkali lye=1: 10W/V) centrifugal behind microwave treatment 15min under the 480W power, it is neutral that washing precipitation to supernatant is for 20% sodium hydroxide solution of water=1: 1; Get above-mentioned deposition, add 5% acetic acid soln and under 100 ℃, handle 5h, spinning, regulating the pH value is 8～10, and a large amount of white flockss appear in this moment, and are centrifugal, and it is neutral that vaal water washing precipitation to supernatant is; Transfer is precipitated in the plate, 50 ℃ of following vacuum-dryings, and the gained solid is chitosan.
Beneficial effect of the present invention:
1. technology of the present invention is simple, be convenient to produce;
2. productive rate of the present invention is higher.
Description of drawings:
Fig. 1, Fig. 2, Fig. 3 are the influence figures of three factors to the chitosan productive rate.
Embodiment:
The present invention is a raw material with the black mold, utilizes fermentation method to prepare chitosan, and process step of the present invention is:
1. fermentation of Aspergillus niger
The slant medium preparation: 4% agar, 2% yeast extract paste, 2% peptone, 4% sucrose, the test tube of packing into after the heating and filtering is in 121 ℃ of autoclaving 20min; On the aseptic technique platform, inoculate, cultivate 0～110h in 22～34 ℃ of electro-heating standing-temperature cultivators; Scrape the mould spore of the black marrow of getting on the fresh inclined-plane with transfering loop; Change over to and sterilized water is housed and is equipped with in the tool plug triangular flask of granulated glass sphere; Fully suspension concentration is calculated with blood counting chamber in the vibration back, and is diluted to desired concn (108 spores of 7108～9X/mL) with sterilized water; In the 500mL Erlenmeyer flask, encase the Erlenmeyer flask bottleneck, in 121 ℃ of autoclaving 20min with multilayer gauze and kraft paper; Spore suspension is inserted according to quantity at the aseptic technique platform in the sterilization back, the constant temperature shaker fermentation, and shaking speed is 120～210r/min;
2. the preparation of chitosan
Get fermented liquid, centrifuge washing thalline to supernatant is colourless, and (thalline: alkali lye=1: 10w/V), centrifugal behind 50 ℃ of following processing 3h, it is colourless that water washing is precipitated to supernatant to get gained deposition adding 7% sodium hydroxide solution; Get above-mentioned deposition, add ethanol: (thalline: alkali lye=1: 10W/V) centrifugal behind microwave treatment 15min under the 480W power, it is neutral that washing precipitation to supernatant is for 20% sodium hydroxide solution of water=1: 1; Get above-mentioned deposition, add 5% acetic acid soln and under 100 ℃, handle 5h, spinning, regulating the pH value is 8～10, and a large amount of white flockss appear in this moment, and are centrifugal, and it is neutral that vaal water washing precipitation to supernatant is; Transfer is precipitated in the plate, 50 ℃ of following vacuum-dryings, and the gained solid is chitosan.
The present invention selects leavening temperature, fermentation time, shaking speed for influencing the factor of chitosan productive rate, the visible Fig. 1 of the situation that influences, Fig. 2, Fig. 3.
Can be seen that by Fig. 2 bacterial strain is in culturing process, the output of chitosan and relative molecular mass are all changing.Cultivate the primary mycelium ramp, the chitosan amount also increases sharply.Along with the increase of incubation time, the chitosan amount reaches capacity gradually, and mycelial amount no longer increases; On the other hand, the relative molecular mass of chitosan descends rapidly along with the prolongation of during cultivation.
As a kind of preferred plan, preparation process of the present invention is: leavening temperature is 28 ℃, and shaking speed is 180r/min, and fermentation time is 80h.
Claims (2)
1. the preparation method of a chitosan is characterized in that, preparation process of the present invention is:
(1) fermentation of Aspergillus niger
The slant medium preparation: 4% agar, 2% yeast extract paste, 2% peptone, 4% sucrose, the test tube of packing into after the heating and filtering is in 121 ℃ of autoclaving 20min; On the aseptic technique platform, inoculate, cultivate 0～110h in 22～34 ℃ of electro-heating standing-temperature cultivators; Scrape the mould spore of the black marrow of getting on the fresh inclined-plane with transfering loop; Change over to and sterilized water is housed and is equipped with in the tool plug triangular flask of granulated glass sphere; Fully suspension concentration is calculated with blood counting chamber in the vibration back, and is diluted to desired concn (108 spores of 7108～9X/mL) with sterilized water; In the 500mL Erlenmeyer flask, encase the Erlenmeyer flask bottleneck, in 121 ℃ of autoclaving 20min with multilayer gauze and kraft paper; Spore suspension is inserted according to quantity at the aseptic technique platform in the sterilization back, the constant temperature shaker fermentation, and shaking speed is 120～210r/min;
(2) preparation of chitosan
Get fermented liquid, centrifuge washing thalline to supernatant is colourless, and (thalline: alkali lye=1: 10w/V), centrifugal behind 50 ℃ of following processing 3h, it is colourless that water washing is precipitated to supernatant to get gained deposition adding 7% sodium hydroxide solution; Get above-mentioned deposition, add ethanol: (thalline: alkali lye=1: 10W/V) centrifugal behind microwave treatment 15min under the 480W power, it is neutral that washing precipitation to supernatant is for 20% sodium hydroxide solution of water=1: 1; Get above-mentioned deposition, add 5% acetic acid soln and under 100 ℃, handle 5h, spinning, regulating the pH value is 8～10, and a large amount of white flockss appear in this moment, and are centrifugal, and it is neutral that vaal water washing precipitation to supernatant is; Transfer is precipitated in the plate, 50 ℃ of following vacuum-dryings, and the gained solid is chitosan.
2. the preparation method of a kind of chitosan according to claim 1 is characterized in that, preparation process of the present invention is: leavening temperature is 28 ℃, and shaking speed is 180r/min, and fermentation time is 80h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2010105160348A CN102453739A (en) | 2010-10-21 | 2010-10-21 | Preparation method of chitosan |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2010105160348A CN102453739A (en) | 2010-10-21 | 2010-10-21 | Preparation method of chitosan |
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| CN102453739A true CN102453739A (en) | 2012-05-16 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103100107A (en) * | 2013-01-25 | 2013-05-15 | 安吉县阳光医药用品有限责任公司 | Method for preparing medical chitin biological dressing |
| CN103952454A (en) * | 2014-05-04 | 2014-07-30 | 青岛科柏利高性能聚合物有限公司 | Process for preparing degradable plastics by using kitchen wastes |
| CN109724956A (en) * | 2018-12-28 | 2019-05-07 | 广西壮族自治区蚕业技术推广总站 | The detection method and its dyeing liquor and preparation method of lepidopterous insects particulate protozoon spore |
| CN111893152A (en) * | 2020-09-02 | 2020-11-06 | 江西师范大学 | Method for biosynthesizing chitosan by using bacteria |
| CN114395280A (en) * | 2022-02-22 | 2022-04-26 | 盐城市德莫环保科技有限公司 | Improved environment-friendly coating containing diatom ooze for green building and preparation method |
-
2010
- 2010-10-21 CN CN2010105160348A patent/CN102453739A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103100107A (en) * | 2013-01-25 | 2013-05-15 | 安吉县阳光医药用品有限责任公司 | Method for preparing medical chitin biological dressing |
| CN103100107B (en) * | 2013-01-25 | 2014-08-20 | 安吉县阳光医药用品有限责任公司 | Method for preparing medical chitin biological dressing |
| CN103952454A (en) * | 2014-05-04 | 2014-07-30 | 青岛科柏利高性能聚合物有限公司 | Process for preparing degradable plastics by using kitchen wastes |
| CN109724956A (en) * | 2018-12-28 | 2019-05-07 | 广西壮族自治区蚕业技术推广总站 | The detection method and its dyeing liquor and preparation method of lepidopterous insects particulate protozoon spore |
| CN111893152A (en) * | 2020-09-02 | 2020-11-06 | 江西师范大学 | Method for biosynthesizing chitosan by using bacteria |
| CN111893152B (en) * | 2020-09-02 | 2021-04-06 | 江西师范大学 | Method for biosynthesizing chitosan by using bacteria |
| CN114395280A (en) * | 2022-02-22 | 2022-04-26 | 盐城市德莫环保科技有限公司 | Improved environment-friendly coating containing diatom ooze for green building and preparation method |
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Application publication date: 20120516 |