CN109988796A - A kind of fermentation medium and its method for cultivating bacteria cellulose film - Google Patents
A kind of fermentation medium and its method for cultivating bacteria cellulose film Download PDFInfo
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Abstract
The invention discloses a kind of fermentation medium and its method for cultivating bacteria cellulose film, the formula of fermentation medium includes: that carbon source 33g, nitrogen source 24g, inorganic salts 1.5g, sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml is added by every liter of culture medium；The method that the present invention regulates and controls the pH of fermentation medium by the formula cooperation addition acetic acid of culture medium, realizes without sterilizing, can have outstanding fungistatic effect；And adjust pH in 4.0-4.2, there is outstanding absorbent after the cleaned water with pressure of obtained product, improve and use comfort, quickening wound healing；Obtained product is once purged to have outstanding transparency, facilitates observation wound situation, can be used as a visual wound care dressings, has a good application prospect in wound surface dressing.
Description
Technical field
The present invention relates to a kind of bacteria cellulose production field, especially a kind of fermentation medium and its culture bacterial fibers
The method of plain film.
Background technique
Bacteria cellulose (Bacterial Cellulose, abbreviation BC) is a kind of natural fiber generated by bacterial fermentation
The general designation of element.By acetic acid Pseudomonas (Acetobacter), Agrobacterium (Agrobacterium), rhizobium
(Rhizobium) certain microbial fermentation and in Sarcina (Sarcina) etc. generates, and typically representing bacterium is wood Portugal
Sweet and sour acidfast bacilli (Glucoacetobacter xylinum).Bacteria cellulose and plant cellulose chemical group having the same
At, as D- glucopyranose acid anhydride with chain macromolecule made of β-Isosorbide-5-Nitrae-glucosides key connection, but the morphosis of bacteria cellulose
It is very different with supramolecular structure and plant cellulose, belongs to typical biological nano fibrous material.Currently, utilizing micro- life
Object fermented-producing bacteria cellulose has been widely used in terms of food, papermaking, medicine and bioengineering.
During carrying out bacteria cellulose productive culture, as medium sterilization is not thorough, vessel disinfection is not thorough and trains
Microbiological contamination phenomenon may all occur during the cultivation process in the problems such as feeding environmental Kuznets Curves are not stringent.Carrying out bacteria cellulose Membrance cuiture mistake
Cheng Zhong, microbiological contamination are broadly divided into two periods: static culture early period after inoculation, and main contaminated bacteria is saccharomycete, ferment at this stage
Female bacterium can mushroom out breeding, and meeting eccrine fiber element enzyme in the medium, often will cause in bacteria cellulose culture big
The destructive loss of area.In the static culture later period, main contaminated bacteria is mould and bacterium, wherein germ contamination situation at this stage
Seldom occur.Since mould microbiological contamination occurred in the later period, bacteria cellulose film has certain thickness, and the growth and breeding speed of mould
It spends relatively slow, generally only will cause small-scale local losses.
In conventional bacteria cellulose production technology, culture medium generally requires the process by high-temperature sterilization, although high temperature
Sterilizing can kill the microorganism in culture medium, but high temperature can also destroy some nutritional ingredients in culture medium, cause culture medium nutrition
The loss of ingredient, while autoclaving process needs a series of resources such as corollary equipment, time and energy.Therefore, break biography to go out
Bacterium technique, develop it is a kind of be not required to sterilizing culture medium of bacterial cellulose preparation method bacteria cellulose produce fermentation application in not
It is only capable of that the microbiological contamination in bacteria cellulose incubation is inhibited to happen, improves row yielding, needed for capable of also cutting down in sterilization process
Equipment, time and the energy, shorten period and cost.
General wound dressing such as gauze, non-woven fabrics application and foam class dressing etc., due to its material property, transparency is not
Height cannot directly observe wound situation.Situations such as infection or secondary damage occurs such as wound, cannot be by directly observing, in time
It was found that wound situation, certain influence is brought to patient.A kind of material that can be used as visual wound dressing is developed for wound
Surface of a wound covering, situations such as observing wound situation, find patient wound's infection or secondary damage in time, curing to patient wound has very
Good facilitation.
It is a kind of good without the bacteria cellulose film transparency that sterilizes and produce that market needs, and the good bacterium of absorbent is fine
Plain cultural method is tieed up, the present invention solves such problems.
Summary of the invention
To solve the deficiencies in the prior art, the purpose of the present invention is to provide a kind of fermentation medium and its culture bacterium are fine
The method for tieing up plain film is regulated and controled the method for the pH of fermentation medium by the formula cooperation addition acetic acid of culture medium, realized without going out
Bacterium can have outstanding fungistatic effect；And adjust pH in 4.0-4.2, have after the cleaned water with pressure of obtained product
Outstanding absorbent improves and uses comfort, accelerates wound healing；Obtained product is once purged with outstanding transparent
Degree facilitates observation wound situation.
In order to achieve the above objectives, the present invention adopts the following technical scheme that:
A kind of fermentation medium, formula include: that carbon source 33g, nitrogen source 24g, inorganic salts 1.5g, hydrogen is added by every liter of culture medium
Sodium oxide molybdena 2g, citric acid 0.8g and acetic acid 3ml.
A kind of fermentation medium above-mentioned, carbon source include: glucose 20g, sucrose 8g, glycerol 5g.
A kind of fermentation medium above-mentioned, nitrogen source include: peptone 10g, corn pulp 7g, ammonium sulfate 7g.
A kind of fermentation medium above-mentioned, inorganic salts include: sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g, sulfuric acid
Magnesium 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g.
A kind of method of fermentation medium culture bacteria cellulose film, the formula of fermentation medium include: to cultivate by every liter
Carbon source 33g, nitrogen source 24g, inorganic salts 1.5g, sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml is added in base；
Cultural method includes the following steps:
Step 1, according to recipe configuration fermentation medium；
Step 2 adjusts 4.2 or less fermentation medium pH to acid solution；
Step 3 accesses acetobacter xylinum in the fermentation medium for regulating pH；
Step 4 dispenses the culture medium after inoculation after mixing evenly, and breathable sealing, quiet under 28~30 DEG C of environment
State culture 5~8 days, obtain bacteria cellulose film.
A kind of method of fermentation medium culture bacteria cellulose film above-mentioned, acid solution is acetic acid.
A kind of method of fermentation medium culture bacteria cellulose film above-mentioned, step 2 adjust fermented and cultured with acetic acid
Base pH is 4.0~4.2.
A kind of method of fermentation medium culture bacteria cellulose film above-mentioned, step 3, in the fermentation training for regulating pH
It supports in base and accesses acetobacter xylinum, the access amount of acetobacter xylinum accounts for 6~20% (v/v) of fermented and cultured base unit weight.
The invention has the beneficial effects that:
The present invention needs not move through high-temperature sterilization using the method for adjusting Medium's PH Value reaching culture medium and can be inoculated into
The culture of row bacteria cellulose ensure that medium nutrient content is not destroyed, also reduce bacteria cellulose film and cultivating
Microbiological contamination situation in the process, improves row yielding, while reducing high-temperature sterilization link again, reduces culture threshold, deduction and exemption
Resource needed for autoclaving process, such as equipment, the energy, manpower and time, shorten the period, and culture bacterium is greatly saved
The cost of cellulose, can adapt in large-scale production；
Contain acetic acid in culture medium prescription, on the one hand can form buffer salt system with composition some other in culture medium and maintain
Medium pH, another aspect acetic acid can be used as the carbon source and synergistic factor that acetobacter xylinum utilizes, improves the yield of bacteria cellulose；
Obtained bacteria cellulose film using method culture of the invention is once purged, the diaphragm before pressure water and after pressure water
Being overlying on observation on human skin has the transparency well；Meanwhile pressing the diaphragm after water that there is good absorbent, in conjunction with thin
The characteristics of fungin good biocompatibility itself, can be used as a visual wound care dressings, in wound surface dressing
It has a good application prospect.
Specific embodiment
Below in conjunction with detailed description of the invention by specific embodiments.
A kind of fermentation medium, formula include: that carbon source 33g, nitrogen source 24g, inorganic salts 1.5g, hydrogen is added by every liter of culture medium
Sodium oxide molybdena 2g, citric acid 0.8g and acetic acid 3ml.
As a kind of preferred embodiment, carbon source includes: glucose 20g, sucrose 8g, glycerol 5g.
As a kind of preferred embodiment, nitrogen source includes: peptone 10g, corn pulp 7g, ammonium sulfate 7g.
As a kind of preferred embodiment, inorganic salts include: sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g, magnesium sulfate
0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g.
A kind of method of fermentation medium culture bacteria cellulose film, includes the following steps:
Step 1, according to recipe configuration fermentation medium.
Step 2 adjusts 4.2 or less fermentation medium pH to acid solution；As a preference, acid solution is acetic acid.Make
To be a kind of preferred, pH adjustable range is 4.0~4.2.
Step 3 accesses acetobacter xylinum in the fermentation medium for regulating pH；As a preference, acetobacter xylinum connects
Enter 6~20% (v/v) that amount accounts for fermented and cultured base unit weight.
Step 4 dispenses the culture medium after inoculation after mixing evenly, and breathable sealing, quiet under 28~30 DEG C of environment
State culture 5~8 days, obtain bacteria cellulose film.
Below by way of experimental verification beneficial effects of the present invention；
△ experiment one, the verifying present invention can inhibit varied bacteria growing in conjunction with the adjusting of PH by the formula of culture medium；
Experimentation:
Acetobacter xylinum in following examples is purchased from China Committee for Culture Collection of Microorganisms's common micro-organisms center, bacterium
Kind number 1.1812.
Embodiment 1
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It, no microbiological contamination obtains bacteria cellulose diaphragm, and it is 2.8g that dry film weight is measured after cleaning-drying.
Embodiment 2
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 5.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It, whole microbiological contaminations cannot obtain bacteria cellulose diaphragm.
Embodiment 3
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.0 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 20% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 5
It, no microbiological contamination obtains bacteria cellulose diaphragm, and it is 3.8g that dry film weight is measured after cleaning-drying.
Embodiment 4
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.8 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 20% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 5
It, whole microbiological contaminations cannot obtain bacteria cellulose diaphragm.
Embodiment 5
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.5 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 20% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 5
It, local microbiological contamination cannot obtain bacteria cellulose diaphragm.
△ show that acetic acid adjusts culture medium difference pH and microbiological contamination situation by as above experiment, as a result such as table 1:
| The pH of acetic acid adjusting culture medium | Microbiological contamination situation | |
| The product that embodiment 1 obtains | 4.2 | Without microbiological contamination |
| The product that embodiment 2 obtains | 5.2 | Microbiological contamination |
| The product that embodiment 3 obtains | 4.0 | Without microbiological contamination |
| The product that embodiment 4 obtains | 4.8 | Microbiological contamination |
| The product that embodiment 5 obtains | 4.5 | Microbiological contamination |
Interpretation of result: in the case where being lower than 4.2 with the pH that acetic acid adjusts culture medium, the growth of bacterium can be inhibited.
Acetobacter xylinum is used for sweet and sour brewing in food service industry, and Dichlorodiphenyl Acetate and low ph conditions have good adaptability.We
Acetic acid is added in method in the medium, forms lower pH environment, can inhibit miscellaneous bacteria well, after being inoculated with acetobacter xylinum, due to
Its strain properties can quickly be bred, and formed dominant microflora, further suppressed the growth and breeding of other miscellaneous bacterias, reached and do not had to go out
Bacterium can also inhibit the effect of miscellaneous bacteria, and then achieve the purpose that not unpasteurized fermentation medium fermented-producing bacteria cellulose.
Acetic acid can participate in tricarboxylic acid cycle (the Tricarboxylic Acid of acetobacter xylinum as a kind of organic acid
Cycle, TCA), promote metabolic fluxes to turn to TCA circulation from the synthesis of cellulose in the growth early stage of acetobacter xylinum, generates more
Energy accelerates the growth and division of bacterium, produces in conjunction in prior fermentation culture medium almost without bacteria cellulose, and culture solution is sticky
The low movement that will not inhibit acetobacter xylinum is spent, it is more than the quantity of acetobacter xylinum and uniform in early stage fermentation medium.In the wooden vinegar bar
When the growth intermediary and later stages metabolic fluxes of bacterium turn to the synthesis of cellulose from TCA circulation, fiber that acetobacter xylinum synthesizes in the medium
Element distribution is more uniform, avoids diaphragm transparency caused by film surface is reunited because of fiber undue growth and internal fiber and declines,
And diaphragm hole caused by reduction film surface fiber undue growth and the reunion of film internal fiber is small, the low phenomenon of absorbent.
Therefore, the distribution of bacterial fibers diaphragm fiber and hole that this method culture obtains are uniform, have good imbibition rate and transparency.
△ experiment two；
There is good absorbent in order to verify after bacteria cellulose film that this method culture obtains is cleaned and pressure water,
Do following experiment:
Experimentation:
According to following examples culture bacteria cellulose film, water is pressed to calculate imbibition rate after cleaning；
Embodiment 6
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It, no microbiological contamination obtains bacteria cellulose diaphragm.
(5) will culture obtain bacteria cellulose diaphragm cleaning after pressure water to moisture content be 95%, be cut into 10cm ×
10cm weighs initial weight W1It for 11.6g, is put into physiological saline, stands 24 hours, it is raw to suck surface with blotting paper or towel
Salt water is managed, the weight W after weighing 24 hours2For 36.3g, by formula (W2 ˉW1)/W1It is 212.9% that imbibition rate for 24 hours, which is calculated,.
Embodiment 7
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.0 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 20% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 5
It, no microbiological contamination obtains bacteria cellulose diaphragm.
(5) will culture obtain bacteria cellulose diaphragm cleaning after pressure water to moisture content be 95%, be cut into 10cm ×
10cm weighs initial weight W1It for 12.4g, is put into physiological saline, stands 24 hours, it is raw to suck surface with blotting paper or towel
Salt water is managed, the weight W after weighing 24 hours2For 40.6g, by formula (W2 ˉW1)/W1It is 227.4% that imbibition rate for 24 hours, which is calculated,.
Embodiment 8
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
At fermentation medium, 105 DEG C of sterilizing 15min are spare after cooling.
(2) pH is adjusted: adjusting medium pH to 5.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It.
(5) will culture obtain bacteria cellulose diaphragm cleaning after, pressure water to moisture content be 95%, be cut into 10cm ×
10cm weighs initial weight W1It for 10.2g, is put into physiological saline, stands 24 hours, it is raw to suck surface with blotting paper or towel
Salt water is managed, the weight W after weighing 24 hours2For 24.7g, by formula (W2ˉW1)/W1It is 142.2% that imbibition rate for 24 hours, which is calculated,.
By comparing as above experiment, draw the following conclusions.
If regulating and controlling the pH range of fermentation medium in 4.0-4.2 by addition acetic acid, bacteria cellulose diaphragm imbibition rate for 24 hours
Greater than 200%, there is good absorbent after illustrating the cleaned water with pressure of diaphragm that the method for the present invention culture obtains.
△ experiment three:
After the bacteria cellulose Membrane cleaning obtained for verification method culture, the diaphragm before pressure water and after pressure water has transparent
The good feature of property, does following experiment:
According to following examples culture bacteria cellulose film, transparency is observed after cleaning；
Embodiment 9
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It, no microbiological contamination obtains bacteria cellulose diaphragm.
(5) obtained bacteria cellulose diaphragm will be cultivated, is overlying on human skin and observes with the diaphragm after pressure water before pressure water,
Can clear view to skin, diaphragm transparency is good.
Embodiment 10
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
It is spare at fermentation medium.
(2) pH is adjusted: adjusting medium pH to 4.0 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 20% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 5
It, no microbiological contamination obtains bacteria cellulose diaphragm.
(5) after the bacteria cellulose diaphragm cleaning obtained culture, human skin is overlying on the diaphragm after pressure water before pressure water
Upper observation, energy clear view to skin, diaphragm transparency are good.
Embodiment 11
(1) preparation of culture medium: carbon source 33g (glucose 20g, sucrose 8g and glycerol 5g) is added by every liter of culture medium, nitrogen source
24g (peptone 10g, corn pulp 7g, ammonium sulfate 7g), inorganic salts 1.5g (sodium chloride 0.4g, potassium chloride 0.1g, calcium chloride 0.2g,
Magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g), sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml configuration
At fermentation medium, 105 DEG C of sterilizing 15min are spare after cooling.
(2) pH is adjusted: adjusting medium pH to 5.2 with acetic acid.
(3) it is inoculated with: accessing acetobacter xylinum, stirring by the inoculum concentration of 6% (accounting for culture medium) in above-mentioned culture medium.
(4) tray is poured into quantitative 500ml packing, with preservative film puncture breathable sealing, under 28 DEG C of environment, static culture 8
It.
(5) it after the bacteria cellulose diaphragm cleaning obtained culture, is observed before pressing water with the diaphragm after pressure water, diaphragm
Poor transparency.
Show that acetic acid adjusts medium pH and diaphragm transparency situation by as above testing, as a result such as the following table 2:
Table 2
By comparing as above experiment, draw the following conclusions.
If having after 4.0-4.2, bacteria cellulose diaphragm cleaning by the pH range that addition acetic acid regulates and controls fermentation medium
There is outstanding transparency, can be used as visual wound care dressings.
The present invention, which is realized through acetic acid, adjusts non-sterilising medium pH to inhibit in bacteria cellulose incubation
Microbiological contamination happens, and the lower antibacterial situation of culture medium medium pH is better；When adjusting pH is 4.0-4.2, before pressure water and press
Diaphragm after water, which is overlying on observation on human skin, has the transparency well；Meanwhile pressing the diaphragm after water that there is good imbibition
Performance can be used as a visual wound care dressings, in wound in conjunction with the characteristics of bacteria cellulose good biocompatibility itself
It is had a good application prospect on Wound dressing.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the above embodiments do not limit the invention in any form, all obtained by the way of equivalent substitution or equivalent transformation
Technical solution is fallen within the scope of protection of the present invention.
Claims (8)
1. a kind of fermentation medium, which is characterized in that formula includes: to be added carbon source 33g by every liter of culture medium, nitrogen source 24g, inorganic
Salt 1.5g, sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml.
2. a kind of fermentation medium according to claim 1, which is characterized in that the carbon source includes: glucose 20g, sugarcane
Sugared 8g, glycerol 5g.
3. a kind of fermentation medium according to claim 1, which is characterized in that the nitrogen source includes: peptone 10g, beautiful
Rice & peanut milk 7g, ammonium sulfate 7g.
4. a kind of fermentation medium according to claim 1, which is characterized in that the inorganic salts include: sodium chloride 0.4g,
Potassium chloride 0.1g, calcium chloride 0.2g, magnesium sulfate 0.1g, sodium dihydrogen phosphate 0.3g, potassium dihydrogen phosphate 0.4g.
5. a kind of method of fermentation medium culture bacteria cellulose film, which is characterized in that the formula of fermentation medium include: by
Carbon source 33g, nitrogen source 24g, inorganic salts 1.5g, sodium hydroxide 2g, citric acid 0.8g and acetic acid 3ml is added in every liter of culture medium；
Cultural method includes the following steps:
Step 1, according to recipe configuration fermentation medium；
Step 2 adjusts 4.2 or less fermentation medium pH to acid solution；
Step 3 accesses acetobacter xylinum in the fermentation medium for regulating pH；
Step 4 dispenses the culture medium after inoculation after mixing evenly, and breathable sealing, under 28~30 DEG C of environment, static state training
It supports 5~8 days, obtains bacteria cellulose film.
6. a kind of method of fermentation medium culture bacteria cellulose film according to claim 5, which is characterized in that described
Acid solution is acetic acid.
7. a kind of method of fermentation medium culture bacteria cellulose film according to claim 6, which is characterized in that step
Two, adjusting fermentation medium pH with acetic acid is 4.0~4.2.
8. a kind of method of fermentation medium culture bacteria cellulose film according to claim 5, which is characterized in that step
Three, access acetobacter xylinum in the fermentation medium for regulating pH, the access amount of acetobacter xylinum account for fermented and cultured base unit weight 6~
20% (v/v).
Priority Applications (1)
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| CN110964669A (en) * | 2019-12-26 | 2020-04-07 | 青岛科技大学 | Bacterial culture medium containing chitosan, bacterial culture fermentation method and application |
| CN111893152A (en) * | 2020-09-02 | 2020-11-06 | 江西师范大学 | Method for biosynthesizing chitosan by using bacteria |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110964669A (en) * | 2019-12-26 | 2020-04-07 | 青岛科技大学 | Bacterial culture medium containing chitosan, bacterial culture fermentation method and application |
| 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 |
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