CN105296573A - Culture medium for fermentation for recombinant human epidermal growth factor - Google Patents

Culture medium for fermentation for recombinant human epidermal growth factor Download PDF

Info

Publication number
CN105296573A
CN105296573A CN201510843157.5A CN201510843157A CN105296573A CN 105296573 A CN105296573 A CN 105296573A CN 201510843157 A CN201510843157 A CN 201510843157A CN 105296573 A CN105296573 A CN 105296573A
Authority
CN
China
Prior art keywords
culture medium
obtains
ions
distiller
percent
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.)
Pending
Application number
CN201510843157.5A
Other languages
Chinese (zh)
Inventor
韦忠明
黄启斌
莫冬海
马冲
宫猛
包能创
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
HUANUOWEI GENE PHARMAOY CO Ltd GUILIN
Original Assignee
HUANUOWEI GENE PHARMAOY CO Ltd GUILIN
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by HUANUOWEI GENE PHARMAOY CO Ltd GUILIN filed Critical HUANUOWEI GENE PHARMAOY CO Ltd GUILIN
Priority to CN201510843157.5A priority Critical patent/CN105296573A/en
Publication of CN105296573A publication Critical patent/CN105296573A/en
Pending legal-status Critical Current

Links

Landscapes

  • Medicines Containing Plant Substances (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

The invention discloses a culture medium for fermentation for a recombinant human epidermal growth factor. The culture medium comprises the following ingredients by weight: 5.0 to 7.0 percent of glycerin, 3.0 to 3.5 percent of yeast nitrogen base (YNB), 1.8 to 2.2 percent of trace metal ions, 0.5 to 1.0 percent of biotin, 0.5 to 1.0 percent of an accelerating agent, and the balance of water, wherein the trace metal ions are one of iron trichloride ions, calcium chloride ions, sodium molybdate ions, zinc chloride ions, cupric sulfate ions, boric acid ions and aluminum sulfate ions; a phosphate buffer solution is used for adjusting the pH of the culture medium to be 5.5 to 6.5. After the vegetable raw materials are treated with steam explosion, distillation and high-shear mixer treating are carried out to obtain a nano milky vegetable oil ingredient which is high in compatibility to the other ingredients in the culture medium. As the vegetable raw materials are added into the culture medium to serve as the accelerating agent, the yield of a fermentation liquor in the culture medium can be improved by 31 to 42 percent when compared with that in the prior art.

Description

A kind of substratum of recombinant human epidermal growth factor fermentation
Technical field
The present invention relates to biological technical field, particularly relate to the substratum of a kind of recombinant human epidermal growth factor fermentation.
Background technology
Urogastron is a kind of growth regulator found from mouse submandibular gland in the sixties in last century by people such as Cohen, is made up of 53 amino acid, and research confirms that it has the effect stimulated cellular proliferation widely.1975, the people such as Starkey, Cohen were extracted human epidermal growth factor (humanEpidermalGrowthFactor, hEGF) from people's urine, have highly consistent structure, and have consistent biologic activity with M-EGF.Urogastron is extensively present in the various tissue of human body, various kinds of cell can be stimulated to breed, mainly the propagation of epithelial cell and endotheliocyte, and this is by working in conjunction with the EGF-R ELISA (EGFR) on cytolemma.EGF is partly combined outward by the film with cell receptor, activates its tyrosine kinase activity, inducible protein phosphorylation, starts DNA synthesis, activator RNA, protein synthesis, promotes hyperplasia, divides a word with a hyphen at the end of a line.
Urogastron is extensively present in various tissue in human body, and content is atomic, has vital role for the normal physiological function maintaining the various histoorgan of human body (as angle conjunctiva, skin, various mucous membrane, body of gland etc.).With advancing age, body entocuticle growth factor levels declines gradually, show as aging and the degeneration of related tissue's organ, the particularly aging of epithelium, therefore, in time to human body supplementary table skin growth factor, can safeguard human normal function and delay senility, this is the application foundation of Urogastron in beauty and health care industry.In addition, when the impaired grade of human body, endogenous epidermal growth factor can not meet a large amount of needs of tissue repair, in time to damaged part supplementary table skin growth factor, the reparation of impaired when injected organism tissue can be promoted, apply widely clinically, as the corneal epithelial defect reparation caused for a variety of causes such as wound, inflammation, operation, chemical burn and xerophthalmia, the reparation of the various skins such as burn, wound, surgical operation, inflammation, ulcer and Mucosa Defect.
At present, to Urogastron and the existing research of recombinant human epidermal growth factor, such as Chinese patent 97115284.5, disclose epdermal growth favtur (hEGF) engineering bacteria and use the method for this bacterium preparation table skin growth factor, researchist can obtain recombinant human epidermal growth factor (or being called recombinant human epidermal growth factor stoste) by Urogastron.Science and technology in continuous progress, for the research of Urogastron and recombinant human epidermal growth factor also in the middle of constantly carrying out.
Summary of the invention
The object of this invention is to provide the substratum of a kind of recombinant human epidermal growth factor fermentation, this substratum is made up of following part by weight composition:
Described trace metal ion is the one in iron trichloride, calcium chloride, Sodium orthomolybdate, zinc chloride, copper sulfate, boric acid, Tai-Ace S 150;
PH is adjusted to be 5.5-6.5 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, burstpressures 3.0-3.5Mpa, and the dwell time maintains 150-200s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 30-40m/s, time 3-5min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
Preferred further, this substratum is made up of following part by weight composition:
Described trace metal ion is the one in iron trichloride, calcium chloride, Sodium orthomolybdate, zinc chloride, copper sulfate, boric acid, Tai-Ace S 150;
PH is adjusted to be 6.0 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, burstpressures 3.0-3.5Mpa, and the dwell time maintains 150-200s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 30-40m/s, time 3-5min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
Water of the present invention meets pharmaceutical industry standard.
Substratum of the present invention is conventionally prepared.
The usage quantity of Herba Andrographis of the present invention and Japanese Honeysuckle, the volume after preferably pulverizing is equivalent to the 1/4-1/3 of steam explosion tank volume.
Compared with prior art, tool of the present invention has the following advantages:
1, the plant material composition that the present invention uses have passed through steam explosion, distillation, high-shear homogenizing machine process, obtains the plant oil components of nanometer emulsus, and in substratum, the consistency of other formulas is good.
2, the present invention is by adding plant material composition in the medium as promotor, and compared to the prior art, expressing output can improve 31-42%.
Embodiment
With embodiment, the invention will be further described below, but the present invention is not limited to these embodiments.
Embodiment 1:
A substratum for recombinant human epidermal growth factor fermentation, this substratum is made up of following part by weight composition:
Described trace metal ion is iron trichloride;
PH is adjusted to be 5.5 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, and volume is equivalent to 1/3 of steam explosion tank volume, burstpressures 3.5Mpa, and the dwell time maintains 150s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 40m/s, time 3min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
Embodiment 2:
A substratum for recombinant human epidermal growth factor fermentation, this substratum is made up of following part by weight composition:
Described trace metal ion is calcium chloride;
PH is adjusted to be 6.5 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, and volume is equivalent to 1/4 of steam explosion tank volume, burstpressures 3.0Mpa, and the dwell time maintains 200s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 30m/s, time 5min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
Embodiment 3:
A substratum for recombinant human epidermal growth factor fermentation, this substratum is made up of following part by weight composition:
Described trace metal ion is in Sodium orthomolybdate, zinc chloride, copper sulfate, boric acid, Tai-Ace S 150;
PH is adjusted to be 6.0 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, and volume is equivalent to 1/3 of steam explosion tank volume, burstpressures 3.0Mpa, and the dwell time maintains 180s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 35m/s, time 4min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.

Claims (3)

1. a substratum for recombinant human epidermal growth factor fermentation, it is characterized in that, this substratum is made up of following part by weight composition:
Described trace metal ion is the one in iron trichloride, calcium chloride, Sodium orthomolybdate, zinc chloride, copper sulfate, boric acid, Tai-Ace S 150;
PH is adjusted to be 5.5-6.5 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, burstpressures 3.0-3.5Mpa, and the dwell time maintains 150-200s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 30-40m/s, time 3-5min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
2. the substratum of a kind of recombinant human epidermal growth factor fermentation according to claim 1, it is characterized in that, this substratum is made up of following part by weight composition:
Described trace metal ion is the one in iron trichloride, calcium chloride, Sodium orthomolybdate, zinc chloride, copper sulfate, boric acid, Tai-Ace S 150;
PH is adjusted to be 6.0 with phosphate buffered saline buffer;
Described promotor adopts following methods preparation:
1) Feedstock treating: by Herba Andrographis and Japanese Honeysuckle according to etc. heavily mix, clean, drying, pulverize;
2) Steam explosion treatment: by step 1) material that obtains puts into steam explosion tank, burstpressures 3.0-3.5Mpa, and the dwell time maintains 150-200s;
3) distillation process: by step 2) material that obtains puts into distiller, bottom distiller, heating flame or pass into steam, when hot steam is filled in distiller, water vapour, by prolong, is introduced in condenser, again by water-and-oil separator, collect essential oil;
4) high-shear homogenizing machine process: by step 3) essential oil that obtains is by high-shear homogenizing machine process, and linear velocity 30-40m/s, time 3-5min, makes nano-emulsion, and obtains with filtering with microporous membrane is degerming.
3. the substratum of a kind of recombinant human epidermal growth factor fermentation according to claim 1 and 2, it is characterized in that: the usage quantity of described Herba Andrographis and Japanese Honeysuckle, the volume after pulverizing is equivalent to the 1/4-1/3 of steam explosion tank volume.
CN201510843157.5A 2015-11-26 2015-11-26 Culture medium for fermentation for recombinant human epidermal growth factor Pending CN105296573A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510843157.5A CN105296573A (en) 2015-11-26 2015-11-26 Culture medium for fermentation for recombinant human epidermal growth factor

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510843157.5A CN105296573A (en) 2015-11-26 2015-11-26 Culture medium for fermentation for recombinant human epidermal growth factor

Publications (1)

Publication Number Publication Date
CN105296573A true CN105296573A (en) 2016-02-03

Family

ID=55194395

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510843157.5A Pending CN105296573A (en) 2015-11-26 2015-11-26 Culture medium for fermentation for recombinant human epidermal growth factor

Country Status (1)

Country Link
CN (1) CN105296573A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107619070A (en) * 2017-10-13 2018-01-23 桂林融通科技有限公司 Reduce the additive that heavy metal seawater damages to big oyster active oxygen radical
CN107624689A (en) * 2017-10-13 2018-01-26 桂林融通科技有限公司 Reduce the additive that heavy metal seawater influences on big oyster lysozyme activity

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1210145A (en) * 1997-08-29 1999-03-10 中国医学科学院基础医学研究所 Engineering bacteria for epidermal growth factor and preparation of epidermal growth factor by using this bacteria
CN101492675A (en) * 2008-01-21 2009-07-29 吉林圣元科技有限责任公司 Method for producing recombinant human epidermal growth factor acceptor 2 cytoplasmic region with methylotrophy yeast

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1210145A (en) * 1997-08-29 1999-03-10 中国医学科学院基础医学研究所 Engineering bacteria for epidermal growth factor and preparation of epidermal growth factor by using this bacteria
CN101492675A (en) * 2008-01-21 2009-07-29 吉林圣元科技有限责任公司 Method for producing recombinant human epidermal growth factor acceptor 2 cytoplasmic region with methylotrophy yeast

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
JOAN LIN CEREGHINO等: "Heterologous protein expression in the methylotrophic yeast Pichia pastoris", 《FEMS MICROBIOLOGY REVIEWS》 *
崔建云主编: "《食品加工机械与设备》", 30 June 2008, 中国轻工业出版社 *
廖小金等: "重组人表皮生长因子在毕赤酵母中的表达纯化及鉴定", 《厦门大学学报(自然科学版)》 *
王郅媛等: "抗衰老研究新模型—酵母细胞活性氧代谢研究进展", 《食品科学》 *
肖安风等: "毕赤酵母发酵过程胞内活性氧的定量分析", 《泉州师范学院学报(自然科学)》 *
肖安风等: "流式细胞术检测毕赤酵母发酵过程中胞内活性氧水平", 《生物工程学报》 *
陈建烟等: "植物精油生物活性作用机理研究进展", 《天然产物研究与开发》 *
黄秉仁等: "甲基营养型酵母系统表达的重组人表皮生长因子的纯化及其性质", 《中国医学科学院学报》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107619070A (en) * 2017-10-13 2018-01-23 桂林融通科技有限公司 Reduce the additive that heavy metal seawater damages to big oyster active oxygen radical
CN107624689A (en) * 2017-10-13 2018-01-26 桂林融通科技有限公司 Reduce the additive that heavy metal seawater influences on big oyster lysozyme activity

Similar Documents

Publication Publication Date Title
CN105238832A (en) Culture medium for recombined human epidermal growth factor fermentation
CN105296572A (en) Medium used for fermenting recombinant human epidermal growth factor
CN103520082B (en) Anti-aging composition containing epidermal growth factor and preparation method thereof
ATE537814T1 (en) NANOLIPOSOME WITH ESTERIFIED LECITHIN AND PRODUCTION METHOD THEREOF AND COMPOSITION CONTAINING SAME FOR THE PREVENTION OR TREATMENT OF SKIN DISEASES
CN104055700A (en) Anti-crack hand cream and preparation method thereof
CN106619177A (en) Whitening and moisturizing cosmetic of white yak bone collagen protein peptide and preparation method of whitening and moisturizing cosmetic
CN102379838A (en) Method for preparing relaxing and face-cleansing cosmetics and application
CN105296573A (en) Culture medium for fermentation for recombinant human epidermal growth factor
CN1082398A (en) The external-usable hair restorer that contains pine extract
CN105296580A (en) Fermentation medium of recombinant human epidermal growth factor
CN105296576A (en) Fermentation method of recombinant human epidermal growth factor
KR101272792B1 (en) Moisturizing cream with high water retention effect
TW201701868A (en) Personal cleansing and preparation method
CN107029115A (en) A kind of natural hair-loss preventing and hair-restoring stem of noble dendrobium caul-fat
CN107441479A (en) Marine active peptide/chitin burn ointment for treating scald and preparation method thereof
CN105296577A (en) Culture medium for recombinant human epidermal growth factor fermentation
CN105331660A (en) Preparation method of recombinant human epidermal growth factor stock solution
CN108309840A (en) A kind of profession repair membrane facial mask liquid and preparation method thereof
CN105296581A (en) Fermentation technology for recombinant human epidermal growth factor
CN105296575A (en) Fermentation method for recombinant human epidermal growth factor
CN105255976A (en) Manufacturing process of recombined human epidermal growth factors
WO2021217568A1 (en) Assembled jade-activating material for maintaining beauty and keeping young, and assembling and synthesizing method therefor and use thereof
CN105296578A (en) Fermentation process of recombinant human epidermal growth factor
CN114404444A (en) Polysaccharide composition for promoting aquaporin expression and preparation method and application thereof
CN105274173A (en) Recombinant human epidermal growth factor (hEGF) original liquor production process

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
RJ01 Rejection of invention patent application after publication

Application publication date: 20160203

RJ01 Rejection of invention patent application after publication