CN100590193C - Method for producing Nata de Coco by deep layer and superficial layer static state couple fermentation - Google Patents
Method for producing Nata de Coco by deep layer and superficial layer static state couple fermentation Download PDFInfo
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- CN100590193C CN100590193C CN200710030072A CN200710030072A CN100590193C CN 100590193 C CN100590193 C CN 100590193C CN 200710030072 A CN200710030072 A CN 200710030072A CN 200710030072 A CN200710030072 A CN 200710030072A CN 100590193 C CN100590193 C CN 100590193C
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- 238000000855 fermentation Methods 0.000 title claims abstract description 114
- 230000004151 fermentation Effects 0.000 title claims abstract description 114
- 230000003068 static effect Effects 0.000 title claims abstract description 74
- 238000004519 manufacturing process Methods 0.000 title abstract description 9
- 241001602876 Nata Species 0.000 title description 2
- 235000013162 Cocos nucifera Nutrition 0.000 claims abstract description 46
- 244000060011 Cocos nucifera Species 0.000 claims abstract description 45
- 239000007788 liquid Substances 0.000 claims abstract description 42
- 238000000034 method Methods 0.000 claims abstract description 42
- 230000008569 process Effects 0.000 claims abstract description 24
- 108010025899 gelatin film Proteins 0.000 claims abstract description 9
- 230000001954 sterilising effect Effects 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- 238000004659 sterilization and disinfection Methods 0.000 claims description 16
- 238000012262 fermentative production Methods 0.000 claims description 13
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- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 10
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- 108010059892 Cellulase Proteins 0.000 claims description 6
- 229940106157 cellulase Drugs 0.000 claims description 6
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- 230000007062 hydrolysis Effects 0.000 claims description 3
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- 238000012856 packing Methods 0.000 description 14
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 11
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- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 1
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention discloses coupled deep and shallow layer static fermentation process of producing high fiber coconut fruit. The process includes inoculating pyracetic bacillus to the fermentingculture medium for deep layer static fermentation; and subpacking fermented liquid with wet bacillus concentration of 2.0-3.0 g/l in sterilized shallow containers for shallow layer static fermentation, so as to obtain high fiber coconut fruit with gel film in the required thickness. The production process has shortened shallow layer fermentation period, high production efficiency and less hetero bacterial contamination, and is suitable for industrial production.
Description
Technical field
The invention belongs to biological technical field, the method for particularly a kind of deep layer and superficial layer static state couple fermentative production high fibrous coconut.
Background technology
Acetic bacteria is static fermentation in the nutrient solution that with the Sucus Cocois is main raw material, and the gel membranoid substance that fluid surface forms rich cellulose is called high fibrous coconut, and English name is " nata de coco ".High fibrous coconut is rich in bacteria cellulose, has very high retentiveness and elasticity, and mouthfeel is smooth, chewiness is good, can be used as a kind of good fiber diet, have nourishing functions such as fat-reducing, gut purge stomach, anti-constipation, in very great demand in countries such as China, Japan, Philippines and Indonesia.High fibrous coconut originates in Philippines, and Southeast Asian countries all has production now, and China Hainan Province nearly 100 families of existing high fibrous coconut manufacturer.
High fibrous coconut production is to produce the process that film acetic bacteria aerobic fermentation is produced bacteria cellulose, and domestic and international many researchists have carried out big quantity research to its fermentation condition.Studies show that in the high fibrous coconut fermenting process, air-flow and stirring all can influence the integrity of bacterial cellulose gel film.Adopt deep-layer liquid dynamically ferment (ventilation or stirring etc.), easily form cotton-shaped cenobium in the fermented liquid, the terminal point in submerged fermentation stage is difficult for determining, and each cup fungi bulk concentration is inhomogeneous during packing fermentation tray, thereby cause the fermentation time difference of each fermentation dish bigger, be difficult to be applied to suitability for industrialized production.In order to obtain the cellulosic coconut palm fruit product of high relative molecular weight, and make gel-film complete and not stratified, should adopt tray to carry out shallow-layer liquid static fermentation and produce.But the tray liquid fermenting exists fermentation condition wayward, and floor space is big, and unit surface factory building production capacity is low, and the fermentation period of existing shallow tray fermentation method is longer, shortcomings such as easy pollution microbes.Shorten the shallow-layer liquid static fermentation cycle, improve fermentation efficiency and prevent that living contaminants etc. is still and be badly in need of the key issue that solves in the high fibrous coconut fermentative production.
Summary of the invention
At there being problems such as the shallow-layer liquid static fermentation cycle is long in the existing high fibrous coconut fermentative production, the object of the present invention is to provide the method for a kind of deep layer and superficial layer static state couple fermentative production high fibrous coconut, this method has shortened the shallow-layer liquid static fermentation cycle, improve the fermentation efficiency of unit surface factory building, and reduce the living contaminants chance.
Purpose of the present invention is achieved through the following technical solutions: the method for a kind of deep layer and superficial layer static state couple fermentative production high fibrous coconut comprises the steps:
(1) deep layer static fermentation
With fermention medium process heat sterilization and after being cooled to 28～32 ℃, insert acetobacter xylinum, described acetobacter xylinum inoculum size is 0.025～10.0% (volume percent), in fermentor tank, carry out the deep layer static fermentation, during the fermentation, the fermentation liquid level the space feed sterile air continuously, and control leavening temperature be 28～32 ℃, deep layer static fermentation terminal point is that the wet thallus concentration of fermented liquid reaches 2.0～3.0g/L.
(2) superficial layer static state fermentation
With the wet thallus concentration that obtains in the step (1) is that the fermented liquid branch of 2.0～3.0g/L installs to through in the shallow-layer container of sterilising treatment, carry out the superficial layer static state fermentation, controlled temperature is 26～34 ℃ in the fermenting process, gel-film thickness in the shallow-layer container reaches product specification thickness (to be finished fermentation, promptly obtains high fibrous coconut when thickness is 3～24mm).
In order to realize the present invention better, comprise Sucus Cocois (coconut broken shell water intaking) in the fermention medium in the step (1), the volume percent of described Sucus Cocois is 20～100%; Need regulate pH when described fermention medium is prepared is 3.5～5.5.
The feeding amount of sterile air is that (0.01 * fermentation volume) more than the L/min, feeds sterile air continuously and make the fermentor tank internal pressure reach 0.01～0.02MPa (gauge pressure), in case living contaminants in the step (1).
Step (1) the deep layer static fermentation time is 84～132h, and concrete time and inoculum size, terminal point wet thallus concentration are relevant.
Wet thallus concentration adopts centrifugal weighting method to measure in the step (2): and adding 0.1g cellulase in the 100mL fermented liquid (＞15 υ/mg), 50 ℃ of vibrations of constant temperature 2h, fully after the hydrolysis, replenish distilled water, make volume to 100mL, obtain hydrolyzed solution, then with the 10mL hydrolyzed solution in the centrifugal 20min of 4000r/min, throw out is weighed, and calculate throw out concentration G
1(g/L); Simultaneously, (＞15 υ/mg) be added in the 100mL distilled water mix it through vibration, in kind determine throw out concentration G to take by weighing the 0.1g cellulase
2(g/L); At last, can calculate wet thallus concentration and be (G
1-G
2) (g/L).
The dress liquid height of step (2) middle-shallow layer container is below the 30mm, and specifically adorning the liquid height needs to decide according to product specification.Preferred especially, step (2) middle-shallow layer static fermentation time 40～192h, then the gel-film thickness in the shallow-layer container reaches product specification thickness, and the concrete time needs to decide according to product specification, and gel-film thickness is bigger, and the superficial layer static state fermentation period is longer.
Have according to fermentor tank that floor space is little, fermentation condition is easy to control, be difficult for advantages such as pollution microbes, the present invention gets up fermentor tank and shallow-layer container combination dexterously, has adopted the coupling process of deep-layer liquid static fermentation and shallow-layer liquid static fermentation to produce high fibrous coconut.
The present invention compared with prior art has following advantage and beneficial effect:
(1) the present invention dynamically ferment with deep-layer liquid (ventilation or stirring etc.) compare, the present invention adopts static fermentation technology in the fermentor tank submerged fermentation stage, solved the problem of fermenting and existing dynamically, its fermentation termination is that the wet thallus concentration of fermented liquid reaches 2.0～3.0g/L, and do not need in the fermenting process in fermented liquid, to ventilate and stir, energy consumption is low, and is simple to operate.
(2) the present invention compares with existing superficial layer static state fermentation method, when the shallow-layer container is identical with dress liquid height, the initial cell concentration of middle-shallow layer static fermentation of the present invention is bigger, and cell has been in the metabolizing bacteria Mierocrystalline cellulose stage, time that the superficial layer static state fermentation stage forms gel-film early, thereby the fermentation period that generates the same thickness gel-film is shorter, and its superficial layer static state fermentation period has shortened 20～40% (products of different specifications shortens the amplitude difference); It is remarkable that the superficial layer static state fermentation period shortens amplitude, and the utilization ratio of superficial layer static state fermentation equipment is increased substantially, and can significantly improve the production capacity of unit space fermenting house; In deep layer static fermentation process, be difficult for, and the superficial layer static state fermentation period obviously shortens, and helps reducing the microbiological contamination rate of high fibrous coconut fermentative production, thereby help improving the product yield, reduce production costs by living contaminants.
Description of drawings
Fig. 1 is a process flow diagram of the present invention.
Embodiment
The present invention is described in further detail below in conjunction with embodiment and accompanying drawing, but embodiments of the present invention are not limited thereto.
Embodiment 1
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812 (purchasing institute of microbiology) in the Chinese Academy of Sciences
(1) seed enlarged culturing
Substratum preparation: sucrose 2g, yeast extract paste 0.4g, NH
4Cl 0.4g, MgSO
47H
2O 0.02g, CaCl
20.02g, NaAC 0.01g, FeSO
40.5mg, Sucus Cocois 20mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), is settled to 100mL, regulates pH5.0 with tap water, the 500mL triangular flask of packing into, with 8 layers of gauze wrapping bottleneck, 121 ℃ of sterilization 20min are cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 12h, obtain seed liquor.
(2) deep layer static fermentation
Substratum preparation: sucrose 3000g, yeast extract paste 600g, NH
4Cl 600g, MgSO
47H
2O 30g, CaCl
230g, NaAC 15g, FeSO
40.75g, Sucus Cocois 30L (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 150L with tap water, regulate pH3.5, the 200L fermentor tank of packing into, 121 ℃ of sterilization 20min, insert above-mentioned triangular flask after being cooled to 28～32 ℃ and cultivate sophisticated 100mL seed liquor (inoculum size is 0.067%), carry out the deep layer static fermentation.In the fermenting process, feed sterile air continuously in the space of fermentation liquid level, the sterile air flow is about 4L/min, and making the fermentor tank internal pressure is 0.01～0.02MPa (gauge pressure), in case living contaminants; Simultaneously, temperature is controlled to be 28～32 ℃.Fermentation 108h, wet thallus concentration reaches 2.1g/L.
Wet thallus concentration adopts centrifugal weighting method to measure: get fermented liquid 100mL and put into the 250mL triangular flask, (＞15 υ/mg), 50 ℃ of vibrations of constant temperature 2h is fully after the hydrolysis to add the 0.1g cellulase, replenish distilled water, make volume return to 100mL, accurately draw the 10mL hydrolyzed solution then and put into centrifuge tube, in the centrifugal 20min of 4000r/min, abandoning supernatant, blot the centrifugal tube wall globule with filter paper, throw out is weighed, and calculate throw out concentration G
1(g/L); Simultaneously, (＞15 υ/mg) be added in the 100mL distilled water mix it through vibration, in kind determine throw out concentration G to take by weighing the 0.1g cellulase
2(g/L); At last, can calculate wet thallus concentration and be (G
1-G
2) (g/L).
(3) superficial layer static state fermentation
Install to through the shallow-layer container fermentation dish of sterilising treatment above-mentioned deep layer static fermentation liquid branch (among length=300mm * 220mm * 65mm), dress liquid height is 8mm, cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, place static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 40h can get the high fibrous coconut that thickness is 3mm.
Embodiment 2
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812
(1) seed enlarged culturing
Substratum preparation: sucrose 5g, yeast extract paste 0.3g, NH
4Cl 0.8g, MgSO
47H
2O 0.04g, CaCl
20.04g, NaAC 0.02g, FeSO
41.0mg, Sucus Cocois 80mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), is settled to 200mL, regulates pH5.5 with tap water, the 1000mL triangular flask of packing into, with 8 layers of gauze wrapping bottleneck, 121 ℃ of sterilization 20min are cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 16h, obtain seed liquor.
(2) deep layer static fermentation
Substratum preparation: sucrose 20kg, yeast extract paste 1.2kg, NH
4Cl 3.2kg, MgSO
47H
2O 160g, CaCl
2160g, NaAC 80g, FeSO
44g, Sucus Cocois 320L (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 799.8L with tap water, regulate pH5.5, the 1000L fermentor tank of packing into, 121 ℃ of sterilization 20min, insert above-mentioned triangular flask after being cooled to 28～32 ℃ and cultivate sophisticated 200mL seed liquor (inoculum size is 0.025%), carry out the deep layer static fermentation.In the fermenting process, feed sterile air continuously in the space of fermentation liquid level, the sterile air flow is about 20L/min, and making the fermentor tank internal pressure is 0.01～0.02MPa (gauge pressure), in case living contaminants; Simultaneously, temperature is controlled to be 28～32 ℃.Fermentation 132h, wet thallus concentration reaches 3.0g/L.
(3) superficial layer static state fermentation
Install to through the shallow-layer container fermentation dish of sterilising treatment above-mentioned deep layer static fermentation liquid branch (among length=300mm * 220mm * 65mm), dress liquid height is 8mm, cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, place static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 52h can get the high fibrous coconut that thickness is 5mm.
Embodiment 3
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812
(1) seed enlarged culturing
Seed culture adopts the enlarged culturing flow process of " inclined-plane → triangular flask → seeding tank ", and is specific as follows:
(a) substratum preparation: NH
4Cl 0.4g, MgSO
47H
2O 0.02g, CaCl
20.02g, Sucus Cocois 100mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), regulate pH5.5, the 500mL triangular flask of packing into is with 8 layers of gauze wrapping bottleneck, 121 ℃ of sterilization 20min, be cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 12h, obtain seed liquor.
(b) substratum preparation: NH
4Cl 30g, MgSO
47H
2O 1.5g, CaCl
21.5g, Sucus Cocois 7.5L (the fresh coconut of Wenchang, Hainan, broken shell water intaking), regulate pH5.0, the 10L fermentor tank (seeding tank) of packing into, 121 ℃ of sterilization 20min, insert the sophisticated 100mL seed liquor of step (a) intermediate cam flask culture after being cooled to 28～32 ℃, carry out aeration-agitation and cultivate.In the culturing process, temperature is controlled to be 28～30 ℃, and mixing speed is controlled to be 300r/min, and air flow is controlled to be 0.15～0.25vvm, cultivates 12h, obtains seed liquor.
(2) deep layer static fermentation
Substratum preparation: NH
4Cl 2.8kg, MgSO
47H
2O 140g, CaCl
2140g, Sucus Cocois 700L (the fresh coconut of Wenchang, Hainan, broken shell water intaking), regulate pH4.5, the 1000L fermentor tank of packing into, 121 ℃ of sterilization 20min, insert the sophisticated 7.5L seed liquor of seed tank culture (inoculum size is 1.06%) in the step (b) after being cooled to 28～32 ℃, carry out the deep layer static fermentation.In the fermenting process, feed sterile air continuously in the space of fermentation liquid level, the sterile air flow is about 20L/min, and making the fermentor tank internal pressure is 0.01～0.02MPa (gauge pressure), in case living contaminants; Simultaneously, temperature is controlled to be 28～32 ℃.Fermentation 124h, wet thallus concentration reaches 3.0g/L.
(3) superficial layer static state fermentation
Install to through the shallow-layer container fermentation dish of sterilising treatment above-mentioned deep layer static fermentation liquid branch (among length=300mm * 220mm * 65mm), dress liquid height is 14mm, cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, place static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 84h can get the high fibrous coconut that thickness is 10mm.
Embodiment 4
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812
(1) seed enlarged culturing
Substratum preparation: sucrose 50g, NH
4Cl 8g, MgSO
47H
2O 0.8g, CaCl
20.8g, NaAC0.2g, Sucus Cocois 1600mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 2000mL with tap water, regulate pH5.0, divide to install in 10 1000mL triangular flasks, the bottled 200mL substratum of each triangle, with 8 layers of gauze wrapping bottleneck, 121 ℃ of sterilization 20min are cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 16h, obtain seed liquor.
(2) deep layer static fermentation
Substratum preparation: sucrose 20kg, NH
4Cl 3.2kg, MgSO
47H
2O 160g, CaCl
2160g, NaAC 80g, Sucus Cocois 638L (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), is settled to 798L, regulates pH5.0 with tap water, the 1000L fermentor tank of packing into, 121 ℃ of sterilization 20min insert above-mentioned triangular flask after being cooled to 28～32 ℃ and cultivate sophisticated 2000mL seed liquor (inoculum size is 0.25%), carry out the deep layer static fermentation.In the fermenting process, feed sterile air continuously in the space of fermentation liquid level, the sterile air flow is about 2L/min, and making the fermentor tank internal pressure is 0.01～0.02MPa (gauge pressure), in case living contaminants; Simultaneously, temperature is controlled to be 28～32 ℃.Fermentation 100h, wet thallus concentration reaches 2.2g/L.
(3) superficial layer static state fermentation
Install to through the shallow-layer container fermentation dish of sterilising treatment above-mentioned deep layer static fermentation liquid branch (among length=300mm * 220mm * 65mm), dress liquid height is 19mm, cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, place static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 120h can get the high fibrous coconut that thickness is 15mm.
Embodiment 5
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812
(1) seed enlarged culturing
Seed culture adopts the enlarged culturing flow process of " inclined-plane → triangular flask → seeding tank ", and is specific as follows:
(a) substratum preparation: sucrose 14g, yeast extract paste 0.1g, NH
4Cl 0.8g, MgSO
47H
2O 0.04g, CaCl
20.04g, NaAC 0.02g, FeSO
41.0mg, Sucus Cocois 100mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), is settled to 200mL, regulates pH5.5 with tap water, the 1000mL triangular flask of packing into, with 8 layers of gauze wrapping bottleneck, 121 ℃ of sterilization 20min are cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 16h, obtain seed liquor.
(b) substratum preparation: sucrose 5586g, yeast extract paste 40g, NH
4Cl 320g, MgSO
47H
2O16g, CaCl
216g, NaAC 8g, FeSO
40.4g, Sucus Cocois 39.9L (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 79.8L with tap water, regulate pH5.5, the 100L fermentor tank of packing into, 121 ℃ of sterilization 20min, insert above-mentioned triangular flask after being cooled to 28～32 ℃ and cultivate sophisticated 200mL seed liquor (inoculum size is 0.25%), carry out aeration-agitation and cultivate.In the culturing process, temperature is controlled to be 30 ℃, and mixing speed is controlled to be 300r/min, and air flow is controlled to be 0.10～0.25vvm, cultivates 12h, obtains seed liquor.
(2) deep layer static fermentation
Substratum preparation: sucrose 50.4kg, yeast extract paste 0.36kg, NH
4Cl 2.88kg, MgSO
47H
2O144g, CaCl
2144g, NaAC 72g, FeSO
43.6g, Sucus Cocois 360L (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 720L with tap water, regulate pH5.5, the 1000L fermentor tank of packing into, 121 ℃ of sterilization 20min, insert the sophisticated 80L seed liquor of above-mentioned seed tank culture (inoculum size is 10%) after being cooled to 28～32 ℃, carry out the deep layer static fermentation.In the fermenting process, feed sterile air continuously in the space of fermentation liquid level, the sterile air flow is about 20L/min, and making the fermentor tank internal pressure is 0.01～0.02MPa (gauge pressure), in case living contaminants; Simultaneously, temperature is controlled to be 28～32 ℃.Fermentation 84h, wet thallus concentration reaches 2.0g/L.
(3) superficial layer static state fermentation
Install to through the shallow-layer container fermentation dish of sterilising treatment above-mentioned deep layer static fermentation liquid branch (among length=300mm * 220mm * 65mm), dress liquid height is 30mm, cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, place static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 192h can get the high fibrous coconut that thickness is 24mm.
Comparative example: existing superficial layer static state fermentation method
Bacterial classification: acetobacter xylinum Acetobacter Xylimum 1.1812
(1) seed enlarged culturing
Substratum preparation: sucrose 14g, yeast extract paste 0.1g, NH
4Cl 0.8g, MgSO
47H
2O 0.04g, CaCl
20.04g, NaAC 0.02g, FeSO
41.0mg, Sucus Cocois 100mL (the fresh coconut of Wenchang, Hainan, broken shell water intaking) is settled to 200mL with tap water, regulate pH5.0, the 1000mL triangular flask of packing into is with 8 layers of gauzes wrapping bottleneck, 121 ℃ of sterilization 20min, be cooled to and insert the good slant strains of a ring activation (with conventional transfering loop inoculation) after 28～32 ℃, place 30 ℃ of constant temperature on the shaking culture case, 150r/min shaking culture 12h, place 30 ℃ of static cultivation 24h of incubator constant temperature then, obtain seed liquor.
(2) superficial layer static state is cultivated
Substratum preparation: sucrose 125g, yeast extract paste 0.9g, NH
4Cl 7.1g, MgSO
47H
2O 0.36g, CaCl
20.36g, NaAC 0.18g, FeSO
48.9mg, Sucus Cocois 890mL (the fresh coconut of Wenchang, Hainan, the broken shell water intaking), be settled to 1780mL with tap water, regulate pH4.0,121 ℃ of sterilization 20min, the fermentation dish of packing into aseptic while hot (length=300mm * 220mm * 65mm), cover fermentation Pan Kou and tie with aseptic, gas-pervious fungi-proof paper film, insert above-mentioned triangular flask after being cooled to 26～34 ℃ and cultivate sophisticated 200mL seed liquor (inoculum size is 10.1%), inoculation secondary fermentation dish dress liquid height is 30mm, places static fermentation in the clean fermenting house.In the fermenting process, control fermenting house temperature is 26～34 ℃, and fermentation 288h can get the high fibrous coconut that thickness is 24mm.
Present embodiment and embodiment 5 contrasts illustrate and adopt method of the present invention that the shallow-layer fermentation period has shortened 96h, and the shortening amplitude is 33.3%.
Claims (6)
1, the method for a kind of deep layer and superficial layer static state couple fermentative production high fibrous coconut is characterized in that comprising the steps:
(1) deep layer static fermentation
With fermention medium process heat sterilization and after being cooled to 28～32 ℃, insert acetobacter xylinum, described acetobacter xylinum inoculum size is a volume percent 0.025～10.0%, in fermentor tank, carry out the deep layer static fermentation, during the fermentation, the fermentation liquid level the space feed sterile air continuously, and control leavening temperature be 28～32 ℃, deep layer static fermentation terminal point is that the wet thallus concentration of fermented liquid reaches 2.0～3.0g/L;
(2) superficial layer static state fermentation
With the wet thallus concentration that obtains in the step (1) is that the fermented liquid branch of 2.0～3.0g/L installs to through in the shallow-layer container of sterilising treatment, carry out the superficial layer static state fermentation, controlled temperature is 26～34 ℃ in the fermenting process, finish fermentation when the gel-film thickness in the shallow-layer container reaches 3～24mm, promptly obtain high fibrous coconut;
Comprise Sucus Cocois in the fermention medium in the described step (1), the volume percent of described Sucus Cocois is 20～100%; Regulating pH when described fermention medium is prepared is 3.5～5.5.
2, the method for a kind of deep layer according to claim 1 and superficial layer static state couple fermentative production high fibrous coconut is characterized in that: feed sterile air in the described step (1) continuously and make the fermentor tank internal pressure reach 0.01～0.02MPa.
3, the method for a kind of deep layer according to claim 1 and superficial layer static state couple fermentative production high fibrous coconut, it is characterized in that: described step (1) the deep layer static fermentation time is 84～132h.
4, the method for a kind of deep layer according to claim 1 and superficial layer static state couple fermentative production high fibrous coconut, it is characterized in that: wet thallus concentration adopts centrifugal weighting method to measure in the described step (2): add the 0.1g cellulase in the 100mL fermented liquid, 50 ℃ of vibrations of constant temperature 2h, fully after the hydrolysis, replenish distilled water, make volume to 100mL, obtain hydrolyzed solution, then with the 10mL hydrolyzed solution in the centrifugal 20min of 4000r/min, throw out is weighed, and calculates throw out concentration G
1G/L; Simultaneously, take by weighing the 0.1g cellulase and be added in the 100mL distilled water, it is mixed, in kind determine throw out concentration G through vibration
2G/L; At last, calculating wet thallus concentration is G
1-G
2
5, the method for a kind of deep layer according to claim 1 and superficial layer static state couple fermentative production high fibrous coconut is characterized in that: the dress liquid height of described step (2) middle-shallow layer container is below the 30mm.
6, the method for a kind of deep layer according to claim 1 and superficial layer static state couple fermentative production high fibrous coconut, it is characterized in that: described step (2) the middle-shallow layer static fermentation time is 40～192h.
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| CN103060425B (en) * | 2013-01-07 | 2014-07-09 | 海南椰国食品有限公司 | Application of colored bio-cellulose in cellulase-producing bacterium activity rapid detection |
| CN103385827B (en) * | 2013-07-11 | 2015-07-22 | 海南奈雅生物技术有限公司 | Production technology of coconut and addictive-free mask adding with nylon mesh lining cloth |
| CN103923865B (en) * | 2014-04-28 | 2016-08-17 | 广东轻工职业技术学院 | Bacteria cellulose producing strains and the method utilizing this strain fermentation bacteria cellulose |
| JP7340729B1 (en) * | 2023-03-29 | 2023-09-07 | フジッコ株式会社 | Manufacturing method of Nata de Coco sheet |
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| 高纤椰果啫喱的研制. 杨益衡.食品工业. 2006 |
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