CN105039477A - Method of utilizing waste livestock hair to prepare peptone and application of peptone in fermentation of butyric acid bacteria - Google Patents
Method of utilizing waste livestock hair to prepare peptone and application of peptone in fermentation of butyric acid bacteria Download PDFInfo
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- CN105039477A CN105039477A CN201510400566.8A CN201510400566A CN105039477A CN 105039477 A CN105039477 A CN 105039477A CN 201510400566 A CN201510400566 A CN 201510400566A CN 105039477 A CN105039477 A CN 105039477A
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Abstract
The invention belongs to the technical field of peptone production. Livestock hair is a substance in which content of protein is quite high but cannot be effectively utilized for a long time, so that waste of resources and economy is caused. Demand of peptone during fermentation production is increased constantly, and shortage of resources can be caused inevitably if only conventional raw materials are used to process and produce peptone products. Therefore, a peptone product which has good indexes is produced by utilizing waste cow hair and pig hair as raw materials and subjecting the raw materials to processes of cleaning, smashing, acid hydrolysis, enzyme hydrolysis, decoloring, concentrating, drying and pulverization. The peptone can replace commonly-used peptone to be applied to fermentation production of butyric acid bacteria.
Description
Technical field
The invention belongs to microbiological culture media production technical field, be specifically related to a kind ofly to utilize discarded pig, ox hair sends out as the method for peptone prepared by raw material, relate to the application at fermentative production Butylic acid bacteria of the peptone products that utilizes the method to produce and this product further.
Technical background
The main component of hair does not have lived hard protein matter, is called Keratin sulfate, accounts for about 95% of whole hair.The amino acid chain that Keratin sulfate is formed by about 20 seed amino acids forms.Comprise Gelucystine, methionine(Met), leucine, α-amino-isovaleric acid, Threonine, phenylalanine, Methionin, Histidine, tryptophane, Isoleucine etc., wherein the content of Gelucystine is the highest.Also containing the trace element of kind more than 20 of having an appointment in hair, mainly comprise zinc, calcium, manganese, cadmium, mercury, selenium, copper, cobalt, iron, titanium, molybdenum, nickel etc.It is the byproduct of butchering that pig, ox hair send out, and their major parts are simply discarded, or sells at a bargain and make fertilizer to peasant, because hair pole is not easy to be decomposed by the microorganisms rotten.Therefore, this not only causes the waste of resource, and causes financial loss and environmental pollution to a certain extent.
Peptone is one of of paramount importance nitrogenous source in fermentation industry.Peptone can be divided into animal protein peptone, vegetable protein peptone, microbial proteinous peptone from raw materials source.Tryptone, meat peptone, bone peptone etc. are all animal protein peptones, and soy peptone etc. be vegetable protein peptone, Yeast protein peptone belongs to microbial proteinous peptone.In the last few years, be that the technology of peptone prepared by raw material and product applies for national patent in succession with above-mentioned, as the application number Chinese patent application that is 201410644041.4 discloses a kind of production technique of ox bone peptone, application number be 201310034191.9 Chinese patent application disclose Yeast protein peptone and preparation method thereof, application number be 201110217180.5 Chinese patent application disclose a kind of soybean protein isolate base material that utilizes and produce the method for soy peptone.But the patent of at present China's also open application sends out peptone products into raw material production with pig, ox hair, market do not have yet related products sell.
Clostridium butyricum (Clostridiumbutyricum) is also known as Butylic acid bacteria.Butylic acid bacteria can tolerate hydrochloric acid in gastric juice and enter enteron aisle, secrete the important nutrient butyric acid (butyric acid) of intestinal mucosa regeneration and reparation and can the intestinal beneficial bacteria growings such as bifidus bacillus be promoted, suppress the harmful intestinal tract bacteria growings such as shigella dysenteriae, recovery intestinal microflora balances, reduce the generation of enterotoxin and the murders by poisoning to intestinal mucosa such as amine, ammonia, indoles, recover intestines immunologic function and normal physiological function.This product has remarkable therapeutic action to rat immunity Ulcerative Colitis, IL-8, TNF-α etc. can be suppressed to cause the excessive unconventionality expression of inflammatory factor, suppress the overexpression of resistive connection intestines IgG antibody, reduce bone-marrow-derived lymphocyte transformation efficiency, improve T lymphocyte transformation rate, correct intestines immunologic derangement, recover intestines immunological tolerance, diminish inflammation, ulcer, effectively treat ulcerative colitis.Butylic acid bacteria produces enzyme and vitamins benefit materials in enteron aisle simultaneously, promotes digesting and assimilating of nutritive substance.Current Butylic acid bacteria product is taken by people or is added in animal-feed the probiotics that can be used as a kind of regulating intestinal canal function to, and its domestic market sales volume reaches tens00000000 yuan.In the industrial fermentation of Butylic acid bacteria is produced, conventional peptone is as nitrogenous source.
Based on above elaboration, the invention provides and a kind ofly send out method into raw material production peptone with discarded pig, ox hair, and the product utilizing the method to produce successfully substitutes and commonly uses peptone and be applied in the fermentative production of Butylic acid bacteria.
Summary of the invention
The invention provides and a kind ofly send out method into raw material production peptone with discarded pig, ox hair, and the product utilizing the method to produce is under equivalent cost conditions, can substitute the peptone commonly used in fermentative production, and have higher viable count and gemma number.The present invention is achieved through the following technical solutions:
(1) pig, ox hair being sent out and be placed in different bucket respectively, is 1: 2-4 by weight ratio, and temperature is that the tap water of 40-50 DEG C soaks 15-30 minute, cleaning and taking-up hair.Repeat said process 2-3 time, until dirt is substantially clean.
(2) clean pig, ox hair are sent out to be placed in temperature be respectively that the convection oven of 50 DEG C-60 DEG C dries 4-8 hour, take out dry hair.Pig hair and ox hair are mixed according to certainweight ratio, concrete ratio is the pig hair of 20-30%, the ox hair of 70-80%.Mixing raw material is pulverized by hair pulverizer, and the hair after pulverizing is at 0.2-0.5cm.
(3) get the pig of 100kg, ox hair sends out and 300-500kg contains the mixing of 8-15% citric acid solution, join in high pressure steam stove, pressure is risen to 0.1Mpa-0.2Mpa, keep pressure equilibrium, temperature is 150-180 DEG C, and boiling 3-6 hour carries out acid hydrolysis;
(4) hydrolyzed solution temperature cooled and control at 45-55 DEG C, after regulating pH to 9.0-10.0 with NaOH, adding the Bacillus licheniformis M-Zyme dry powder (200,000 U/g) of hydrolyzed solution weight 0.1-0.3%, carry out enzymolysis 1-3 hour;
(5), after enzymolysis terminates, adjust pH, to 6.0-7.0, after feed liquid being warming up to 75 DEG C-85 DEG C under constantly stirring, and boils 5-10 minute, by enzyme-deactivating.But adding activated carbon decolorizing, consumption is the 1-2% of Tot Prot in raw material, and the time is 1-1.5 hour;
(6) by enzymolysis solution through Plate Filtration, obtain filtrate, through 0.3 μm ~ 0.5 μm ultrafiltration membrance filter, to remove pathogenic bacterium;
(7) enzymolysis solution will be clarified through double-effect evaporator, at 60 DEG C of-80 DEG C of concentrating under reduced pressure, obtain concentrated solution;
(8) concentrated solution is carried out spray dried into powder form in spray tower, control inlet temperature 160 DEG C-175 DEG C, air outlet temperature 85 DEG C-95 DEG C, obtains product.
By the index of the peptone products prepared by aforesaid method be: phosphoric acid salt, alkalescence are detected as without precipitation, clarification, 2% aqueous solution is transparent, potential of hydrogen is 6.2-6.8, and amino nitrogen content is 2.7-2.8%, and tryptophane is 0.85-0.88%, peptone content is 80.3-82.0%, total nitrogen content is 13.5-14.5%, and water content is 3.6-4.0%, and the ash content of coal is 3.5-3.9%, sodium chloride content is 0.16-0.19%, and the molecular weight distribution of peptone is 700 ~ 2500Da.
Advantage of the present invention: it is waste material in butchering that the pig one, used by the present invention, ox hair send out raw material, and therefore the present invention can farthest utilize Biological resources, the waste reducing Biological resources and the pollution reduced environment; Two is that this raw materials technology is through peracid, enzyme two-step hydrolysis, comparatively different merely through a step acid hydrolysis, be hydrolyzed more thorough, be conducive to the quality and the yield that improve peptone, this research applies citric acid first to carry out acid hydrolysis in hair-hydrolyzation in addition, and the hydrochloric acid commonly using about 20% relative to acid hydrolysis reduces and may bring the injuries such as skin corrosivity to human body operation; Three, each index of product obtained is invented excellent.Through company at 10 tons of scale fermentation tank verification experimental verifications, the peptone that this technique is produced, relative to conventional peptone, under equivalent cost condition, obtains higher viable count respectively and produces gemma number.
Embodiment
Example 1
Pig, ox hair being sent out and be placed in different bucket respectively, is 1: 2 by weight ratio, and temperature is that the tap water of 40 DEG C soaks 15 minutes, cleaning and taking-up hair.Repeat said process 2 times, until dirt is substantially clean.Clean pig, ox hair being sent out and being placed in temperature is respectively that the convection oven of 50 DEG C is dried 4 hours, takes out dry hair.Pig hair and ox hair are mixed according to certainweight ratio, concrete ratio is the pig hair of 20% and the ox hair of 80%.Mixing raw material is pulverized by hair pulverizer, and the hair after pulverizing is at 0.2-0.5cm.Get the pig of 100kg, ox hair sends out and 300kg contains 8% citric acid solution mixing, join in high pressure steam stove, pressure is risen to 0.1Mpa, keep pressure equilibrium, temperature is 150 DEG C, and boiling 3 hours, carries out acid hydrolysis.Hydrolyzed solution temperature cooled and controls at 45 DEG C, after regulating pH to 9.0 with NaOH, adding the Bacillus licheniformis M-Zyme dry powder (200,000 U/g) of hydrolyzed solution weight 0.1%, carry out enzymolysis 1 hour.After enzymolysis terminates, adjust pH to 6.0, after feed liquid being warming up to 75 DEG C under constantly stirring, and boils 5 minutes, by enzyme-deactivating.But adding activated carbon decolorizing, consumption is 1% of Tot Prot in raw material, and the time is 1 hour.By enzymolysis solution through Plate Filtration, obtain filtrate, through 0.3 μm of ultrafiltration membrance filter, to remove pathogenic bacterium.Enzymolysis solution will be clarified through double-effect evaporator, at 60 DEG C of concentrating under reduced pressure, obtain concentrated solution.Concentrated solution is carried out spray dried into powder form in spray tower, and control inlet temperature 160 DEG C, air outlet temperature 85 DEG C, obtains product 53.3kg.
By the index of the peptone products prepared by aforesaid method be: phosphoric acid salt, alkalescence are detected as without precipitation, clarification, 2% aqueous solution is transparent, potential of hydrogen is 6.2, and amino nitrogen content is 2.7%, and tryptophane is 0.85%, peptone Content is greater than 82.0%, total nitrogen content is 14.0%, and water content is 4.0%, and the ash content of coal is 3.5%, sodium chloride content is 0.16%, and the molecular weight distribution of peptone is 900 ~ 2000Da.
Example 2
Pig, ox hair being sent out and be placed in different bucket respectively, is 1: 4 by weight ratio, and temperature is that the tap water of 50 DEG C soaks 30 minutes, cleaning and taking-up hair.Repeat said process 3 times, until dirt is substantially clean.Clean pig, ox hair being sent out and being placed in temperature is respectively that the convection oven of 60 DEG C is dried 8 hours, takes out dry hair.Pig hair and ox hair are mixed according to certainweight ratio, concrete ratio is the pig hair of 30% and the ox hair of 70%.Mixing raw material is pulverized by hair pulverizer, and the hair after pulverizing is at 0.2-0.5cm.Get the pig of 100kg, ox hair sends out and 500kg contains 15% citric acid solution mixing, join in high pressure steam stove, pressure is risen to 0.2Mpa, keep pressure equilibrium, temperature is 180 DEG C, and boiling 6 hours, carries out acid hydrolysis.Hydrolyzed solution temperature cooled and controls at 55 DEG C, after regulating pH to 10.0 with NaOH, adding the Bacillus licheniformis M-Zyme dry powder (200,000 U/g) of hydrolyzed solution weight 0.3%, carry out enzymolysis 3 hours.After enzymolysis terminates, adjust pH to 7.0, after feed liquid being warming up to 85 DEG C under constantly stirring, and boils 10 minutes, by enzyme-deactivating.But adding activated carbon decolorizing, consumption is 2% of Tot Prot in raw material, and the time is 1.5 hours.By enzymolysis solution through Plate Filtration, obtain filtrate, through 0.5 μm of ultrafiltration membrance filter, to remove pathogenic bacterium.Enzymolysis solution will be clarified through double-effect evaporator, at 80 DEG C of concentrating under reduced pressure, obtain concentrated solution.Concentrated solution is carried out spray dried into powder form in spray tower, and control inlet temperature 175 DEG C, air outlet temperature 95 DEG C, obtains product 62.7Kg.
By the index of the peptone products prepared by aforesaid method be: phosphoric acid salt, alkalescence are detected as without precipitation, clarification, 2% aqueous solution is transparent, potential of hydrogen is 6.8, and amino nitrogen content is 2.8%, and tryptophane is 0.85%, peptone Content is greater than 81%, total nitrogen content is 13.5%, and water content is 3.8%, and the ash content of coal is 3.9%, sodium chloride content is 0.19%, and the molecular weight distribution of peptone is 700 ~ 1800Da.
Example 3
Pig, ox hair being sent out and be placed in different bucket respectively, is 1: 4 by weight ratio, and temperature is that the tap water of 50 DEG C soaks 15 minutes, cleaning and taking-up hair.Repeat said process 2 times, until dirt is substantially clean.Clean pig, ox hair being sent out and being placed in temperature is respectively that the convection oven of 50 DEG C is dried 4 hours, takes out dry hair.Pig hair and ox hair are mixed according to certainweight ratio, concrete ratio is pig hair and 75% ox hair of 27%.Mixing raw material is pulverized by hair pulverizer, and the hair after pulverizing is at 0.2-0.5cm.Get the pig of 100kg, ox hair sends out and 400kg contains 10% citric acid solution mixing, join in high pressure steam stove, pressure is risen to 0.2Mpa, keep pressure equilibrium, temperature is 160 DEG C, and boiling 4 hours, carries out acid hydrolysis.Hydrolyzed solution temperature cooled and controls at 50 DEG C, after regulating pH to 9.5 with NaOH, adding the Bacillus licheniformis M-Zyme dry powder (200,000 U/g) of hydrolyzed solution weight 0.2%, carry out enzymolysis 2 hours.After enzymolysis terminates, adjust pH to 6.5, after feed liquid being warming up to 75 DEG C under constantly stirring, and boils 5 minutes, by enzyme-deactivating.But adding activated carbon decolorizing, consumption is 1.5% of Tot Prot in raw material, and the time is 1 hour.By enzymolysis solution through Plate Filtration, obtain filtrate, through 0.3 μm of ultrafiltration membrance filter, to remove pathogenic bacterium.Enzymolysis solution will be clarified through double-effect evaporator, at 70 DEG C of concentrating under reduced pressure, obtain concentrated solution.Concentrated solution is carried out spray dried into powder form in spray tower, and control inlet temperature 175 DEG C, air outlet temperature 95 DEG C, obtains product 54.8kg.
By the index of the peptone products prepared by aforesaid method be: phosphoric acid salt, alkalescence are detected as without precipitation, clarification, 2% aqueous solution is transparent, potential of hydrogen is 6.3, and amino nitrogen content is 2.7%, and tryptophane is 0.88%, peptone is containing being greater than most 80.3%, total nitrogen content is 14.5%, and water content is 3.6%, and the ash content of coal is 3.7%, sodium chloride content is 0.18%, and the molecular weight distribution of peptone is 1200 ~ 2500KDa.
Example 4
By the peptone products prepared by example 1,2,3, the Butylic acid bacteria replacing equivalent cost is commonly used peptone and is carried out liquid fermenting test and obtain viable count (calculating with every milliliter of fermented liquid) respectively for 5.6x10
8, 5.9x10
8, 5.4x10
8, gemma number (calculating with every milliliter of fermented liquid) is for 3.4x10
8, 4.0x10
8, 3.7x10
8.The viable count of conventional peptone is 4.7x10
8, gemma number 3.0x10
8.From above result, the peptone prepared by example 1,2,3 is under equivalent cost conditions, and comparatively conventional peptone all improves on viable count and product spore number.
Claims (4)
1. the invention discloses and utilize discarded pig, ox hair sends out and prepare the method for peptone for raw material and utilize the method can be applied in product in fermentative production Butylic acid bacteria, this preparation method characteristic comprises the steps:
(1) pig, ox hair being sent out and be placed in different bucket respectively, is 1: 2-4 by weight ratio, and temperature is that the tap water of 40-50 DEG C soaks 15-30 minute, cleaning and taking-up hair.Repeat said process 2-3 time, until dirt is substantially clean;
(2) clean pig, ox hair are sent out to be placed in temperature be respectively that the convection oven of 50 DEG C-60 DEG C dries 4-8 hour, take out dry hair.Pig hair and ox hair are mixed according to certainweight ratio, concrete ratio is the pig hair of 20-30%, the ox hair of 70-80%.Mixing raw material is pulverized by hair pulverizer, and the hair after pulverizing is at 0.2-0.5cm;
(3) get the pig of 100kg, ox hair sends out and 300-500kg contains the mixing of 8-15% acidic solution, join in high pressure steam stove, pressure is risen to 0.1Mpa-0.2Mpa, keep pressure equilibrium, temperature is 150 DEG C-180 DEG C, and boiling 3-6 hour carries out acid hydrolysis;
(4) hydrolyzed solution temperature cooled and control at 45-55 DEG C, after regulating pH to 9.0-10.0 with NaOH, adding the enzyme dry powder of hydrolyzed solution weight 0.1-0.3%, carry out enzymolysis 1-3 hour;
(5), after enzymolysis terminates, adjust pH, to 6.0-7.0, after feed liquid being warming up to 75 DEG C-85 DEG C under constantly stirring, and boils 5-10 minute, by enzyme-deactivating.But adding activated carbon decolorizing, consumption is the 1-2% of Tot Prot in raw material, and the time is 1-1.5 hour;
(6) by enzymolysis solution through Plate Filtration, obtain filtrate, through 0.3 μm ~ 0.5 μm ultrafiltration membrance filter, to remove pathogenic bacterium;
(7) enzymolysis solution will be clarified through double-effect evaporator, at 60 DEG C of-80 DEG C of concentrating under reduced pressure, obtain concentrated solution;
(8) concentrated solution is carried out spray dried into powder form in spray tower, control inlet temperature 160 DEG C-175 DEG C, air outlet temperature 85 DEG C-95 DEG C, obtains product.
2. preparation method according to claim 1, is characterized in that, acid hydrolysis and the acid involved by enzymolysis and enzyme are respectively citric acid and Bacillus licheniformis M-Zyme (200,000 U/g).
3. the hair protein peptone that preparation method obtains according to claim 1 and 2.
4. hair protein peptone according to claim 3, is characterized in that, the molecular weight distribution of described peptone is 700 ~ 2500Da, and amino nitrogen content is 2.7-2.8%, and total nitrogen content is 13.5-14.5%, and peptone content is 80.3-82.0%.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106086141A (en) * | 2016-08-11 | 2016-11-09 | 大连豪翔生物酶工程有限公司 | Pilus Sus domestica is the method for raw material production feedstuff oligopeptide |
| CN107417437A (en) * | 2017-09-27 | 2017-12-01 | 界首市明亮种植专业合作社 | The preparation method of one boar hair ferment fertilizer |
| CN114292309A (en) * | 2021-12-24 | 2022-04-08 | 百洋产业投资集团股份有限公司 | Method for producing peptone powder from fish processing by-products |
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| CN101550179A (en) * | 2008-03-31 | 2009-10-07 | 北京市科威华食品工程技术有限公司 | Sesbania peptone and production method thereof |
| CN103088098A (en) * | 2013-01-17 | 2013-05-08 | 安徽华强羽绒有限公司 | Method for producing peptone by taking poultry feather as raw material |
| CN103602653A (en) * | 2013-11-20 | 2014-02-26 | 江南大学 | Keratinase with improved thermal stability and specific activity as well as preparation method and application thereof |
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Patent Citations (3)
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| CN101550179A (en) * | 2008-03-31 | 2009-10-07 | 北京市科威华食品工程技术有限公司 | Sesbania peptone and production method thereof |
| CN103088098A (en) * | 2013-01-17 | 2013-05-08 | 安徽华强羽绒有限公司 | Method for producing peptone by taking poultry feather as raw material |
| CN103602653A (en) * | 2013-11-20 | 2014-02-26 | 江南大学 | Keratinase with improved thermal stability and specific activity as well as preparation method and application thereof |
Non-Patent Citations (1)
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| 刘婕: "羽毛降解菌的筛选、鉴定及产酶条件优化", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106086141A (en) * | 2016-08-11 | 2016-11-09 | 大连豪翔生物酶工程有限公司 | Pilus Sus domestica is the method for raw material production feedstuff oligopeptide |
| CN107417437A (en) * | 2017-09-27 | 2017-12-01 | 界首市明亮种植专业合作社 | The preparation method of one boar hair ferment fertilizer |
| CN114292309A (en) * | 2021-12-24 | 2022-04-08 | 百洋产业投资集团股份有限公司 | Method for producing peptone powder from fish processing by-products |
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