阅读说明：本技术 薯渣生产蛋白饲料方法、薯类淀粉加工废水渣利用方法 (Method for producing protein feed from potato residue and method for utilizing potato starch processing wastewater residue ) 是由 赵海 靳艳玲 方扬 谭力 何开泽 于 2019-10-14 设计创作，主要内容包括：本发明公开一种薯渣生产蛋白饲料方法、薯类淀粉加工废水渣利用方法。针对现有技术利用薯类淀粉加工薯渣生产的蛋白饲料真蛋白含量不高,以及加工废水未达到资源化处置水平的缺陷,本发明提供一种薯渣生产蛋白饲料方法。该方法取薯类淀粉加工中产生的新鲜薯渣,采用分阶段发酵,第一阶段加入液化酶与糖化酶,接种鼠李糖乳杆菌或干酪乳杆菌；第二阶段加入尿素,接种产朊假丝酵母；第三阶段接种酿酒酵母。本发明还提供一种薯类淀粉加工废水渣综合利用方法。通过将利用废水制备发酵菌种子液与上述薯渣生产蛋白饲料方法相结合,该综合利用方法能够与现有薯类淀粉加工工艺直接衔接,以一种经济低廉的手段提高了薯类淀粉加工工艺水准。本发明还提供鼠李糖乳杆菌、干酪乳杆菌、产朊假丝酵母与酿酒酵母在薯类淀粉加工废水渣发酵生产蛋白饲料中的应用。(The invention discloses a method for producing protein feed by using potato residues and a method for utilizing potato starch processing wastewater residues. Aiming at the defects that the protein feed produced by processing potato residues by utilizing potato starch in the prior art is not high in true protein content and the processing wastewater does not reach the recycling treatment level, the invention provides a method for producing the protein feed by utilizing the potato residues. The method comprises the steps of taking fresh potato residues generated in potato starch processing, fermenting by stages, adding liquefying enzyme and saccharifying enzyme in the first stage, and inoculating lactobacillus rhamnosus or lactobacillus casei; in the second stage, adding urea and inoculating candida utilis; and inoculating the saccharomyces cerevisiae in the third stage. The invention also provides a comprehensive utilization method of the waste water slag in potato starch processing. By combining the method for preparing zymophyte seed liquid by using waste water with the method for producing protein feed by using potato residues, the comprehensive utilization method can be directly linked with the existing potato starch processing technology, and the potato starch processing technology level is improved by an economic and low-cost means. The invention also provides application of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae in producing protein feed by fermenting potato starch processing wastewater residues.)

1. The method for producing protein feed by using potato dregs utilizes the waste potato dregs in the processing of microbial fermentation potato starch to produce protein feed, and is characterized in that: taking fresh potato residues generated in potato starch processing, performing staged fermentation, adding a liquefying enzyme and a saccharifying enzyme in the first stage, and inoculating lactobacillus rhamnosus or lactobacillus casei; in the second stage, adding urea and inoculating candida utilis; and inoculating the saccharomyces cerevisiae in the third stage.

2. The method of claim 1, wherein: the first stage fermentation condition is 30-37 ℃ closed culture for 12h, the second stage fermentation condition is 28-32 ℃ culture for 12h, and the third stage fermentation condition is 28-32 ℃ culture for 3-7 d.

3. The method of claim 1, wherein: the adding amount of the liquefying enzyme is 0.8-1 per mill of the weight of the potato residue, the adding amount of the saccharifying enzyme is 4-5 per mill of the weight of the potato residue, and the inoculation amount of the lactobacillus rhamnosus or lactobacillus casei is lactobacillus rhamnosus seed liquid accounting for 4-5 percent of the weight of the potato residue.

4. The method of claim 1, wherein: the adding amount of the urea is 0.8-1.5% of the weight of the potato residue, and the inoculation amount of the candida utilis is 2-4% of the weight of the potato residue.

5. The method of claim 1, wherein: the inoculation amount of the saccharomyces cerevisiae is 2-4% of the weight of the potato residue.

6. The method of claim 1, wherein: the adding amount of the liquefying enzyme is 0.8-1 per mill of the weight of the potato residue, the adding amount of the saccharifying enzyme is 4-5 per mill of the weight of the potato residue, and the inoculation amount of the lactobacillus rhamnosus or lactobacillus casei is lactobacillus rhamnosus seed liquid accounting for 4-5 percent of the weight of the potato residue; the adding amount of the urea is 0.8-1.5% of the weight of the potato residue, and the inoculation amount of the candida utilis is 2-4% of the weight of the potato residue; the inoculation amount of the saccharomyces cerevisiae is 2-4% of the weight of the potato residue.

7. The method for utilizing the potato starch processing wastewater and the potato residue, which is realized by the method for producing the protein feed by utilizing the potato residue as claimed in any one of claims 1 to 6, is characterized in that the method comprises the following steps: the inoculation of the lactobacillus rhamnosus or the lactobacillus casei uses a lactobacillus rhamnosus or lactobacillus casei seed solution prepared by culturing lactobacillus rhamnosus or lactobacillus casei by fresh wastewater generated in potato starch processing, the inoculation of the candida utilis uses a candida utilis seed solution prepared by culturing candida utilis by fresh wastewater generated in potato starch processing, and the inoculation of the saccharomyces cerevisiae uses a saccharomyces cerevisiae seed solution prepared by culturing saccharomyces cerevisiae by fresh wastewater generated in potato starch processing.

8. The method for utilizing potato starch processing wastewater slag according to claim 7, characterized in that:

the preparation method of the lactobacillus rhamnosus seed liquid comprises the following steps: boiling fresh wastewater generated in potato starch processing for sterilization, sealing, naturally cooling, inoculating lactobacillus rhamnosus, and culturing at 35-37 ℃ for 24 h;

preparing the lactobacillus casei seed liquid: boiling fresh wastewater generated in potato starch processing for sterilization, sealing, naturally cooling, inoculating lactobacillus casei, and culturing at 35-37 ℃ for 24 h;

the preparation method of the candida utilis seed liquid comprises the following steps: boiling fresh wastewater generated in potato starch processing for sterilization, sealing, naturally cooling, inoculating candida utilis, and culturing at 28-30 ℃ for 36 h;

the preparation of the saccharomyces cerevisiae seed liquid comprises the following steps: boiling fresh wastewater generated in potato starch processing for sterilization, sealing, naturally cooling, inoculating saccharomyces cerevisiae, and culturing at 30-32 ℃ for 48 h.

9. The application of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae in producing protein feed by fermenting potato starch processing waste water residue is characterized in that: firstly, adding liquefying enzyme and saccharifying enzyme into fresh potato residues generated in potato starch processing, inoculating lactobacillus rhamnosus or lactobacillus casei, and carrying out closed fermentation at 30-37 ℃ for 12 h; adding urea, inoculating candida utilis, and fermenting for 12 hours at the temperature of 28-32 ℃; finally inoculating saccharomyces cerevisiae, and fermenting for 3-7 days at 28-32 ℃.

10. The method for producing protein feed by potato pulp as claimed in any one of claims 1 to 6, the method for utilizing potato starch processing wastewater slag as claimed in claim 8, and the application of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae as claimed in claim 9 in the production of protein feed by fermentation of potato starch processing wastewater slag, wherein the method comprises the following steps: the lactobacillus rhamnosus is lactobacillus rhamnosus CICC 20258, the lactobacillus casei is lactobacillus casei CICC 23184, the candida utilis is candida utilis CICC 1314, and the saccharomyces cerevisiae is saccharomyces cerevisiae CCTCC M206111.

Technical Field

The invention relates to a method for producing protein feed, in particular to a method for producing protein feed by using waste residues and waste water generated in the potato starch processing process, belonging to the technical fields of production methods of feed specially suitable for animals, microbial fermentation engineering and resource utilization of food processing waste materials.

Background

A process for producing starch by utilizing root tuber and tuber of potato crops belongs to primary processing production of food, and main byproducts of the process comprise waste residue (namely potato residue) and waste water. At present, the potato starch processing technology mostly adopts the traditional technology, the water consumption is large, and the comprehensive utilization rate of byproducts is low. For example, about 1/6 tons of vermicelli can be obtained per 1 ton of fresh potatoes, and about 4 tons of waste water and 2-3 tons of potato residues are generated at the same time. The main components of the potato residue are starch, cellulose, pectin, protein and the like, and the mass of the potato residue accounts for 45-60% of the mass of the fresh residue according to different production processes. Therefore, the potato residue has higher viscosity. The moisture in the residue is not only tightly bound to the fibers and pectin, but also the undamaged cells can absorb the moisture through the cell membrane, making the removal of the moisture difficult. Similar to potato dregs, the waste water also belongs to high-concentration organic waste water. Although the waste water slag generated in the potato starch processing does not contain toxic and harmful chemical substances, the waste water slag contains a large amount of organic substances, so that the organic pollution to the environment is easily caused. Because the potato starch processing is generally finished by taking fresh potato blocks as raw materials within 3 months of potato harvest, the generation of the waste water residue has the characteristics of obvious concentrated generation time period and huge generation amount in the concentrated time period. In addition, potato starch processing enterprises generally have low starting points, weak strength and low profits, and have no capacity to build or maintain waste water and waste residue treatment facilities which consume money, energy and manpower and do not produce, so that waste residues are randomly discarded to cause rural non-point source environmental pollution, and waste water is directly discharged into natural water without effective treatment, thereby further aggravating the pollution.

In rural areas of potato starch processing sites, farmers often directly feed the starch processing residues as feed to livestock to reduce feed cost. However, the potato residue has low protein content and poor palatability, so that the feed value is not high. With the continuous integration of biotechnology and feed processing industry, the preparation of protein feed for animal feed from potato residue by microbial fermentation is an approach for the resource utilization of potato residue. The currently used microbial species include yeast, mold, bacillus, lactic acid bacteria, actinomycetes, algae, and the like. In order to improve the recycling efficiency, the prior art has two kinds of concepts, one is to screen single strains with higher conversion efficiency (such as long-lasting, Dingfan, Yujinlong and the like, research on biochemical conversion technology of sweet potatoes [ J ]. Chinese food and oil academy, 2017,32(9)), and the other is to adopt compound strain fermentation (such as the Senchen, Dexpension, Wangting and the like, process optimization of producing white feed by fermenting sweet potato residues with microorganisms and influence on the production performance of fattening pigs [ J ]. Jiangsu agricultural science, 2016,44(4), Anvingtin, Sun Shouxin, Liu tree and the like, research on the value-added effect of solid state fermentation sweet potato residues [ J ]. feed research, 2015, 10). The treatment of the waste water mainly focuses on the treatment process of the organic waste water in the environmental pollution treatment, and the flocculant (CN 108328715A, CN110218239A), the coagulating sedimentation and fungal fermentation combined method (CN 109052857A) and the like are adopted to realize the discharge reaching the standard and do not reach the level of the resource treatment like the potato residue.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, provides a method for producing protein feed for animal feed by fermenting potato residues with microorganisms, has low cost and simple process, and can be popularized and applied in rural areas where potato starch is processed. The invention also aims to improve the resource utilization level of the potato starch processing wastewater, realize the comprehensive utilization of the wastewater slag and completely solve the problem of environmental pollution of potato starch processing enterprises in a low-cost manner.

In order to realize the purpose, the invention firstly provides a method for producing protein feed by using potato dregs, and the technical scheme is as follows:

a method for producing protein feed by potato dregs, which utilizes waste potato dregs in the processing of microbial fermentation potato starch to produce protein feed, and is characterized in that: taking fresh potato residues generated in potato starch processing, performing staged fermentation, adding a liquefying enzyme and a saccharifying enzyme in the first stage, and inoculating lactobacillus rhamnosus or lactobacillus casei; in the second stage, adding urea and inoculating candida utilis; and inoculating the saccharomyces cerevisiae in the third stage.

The method for producing the protein feed by using the potato residue utilizes four microorganisms to complete the fermentation process of the potato residue, the potato residue is fresh potato residue produced from a potato starch processing line, and the water content is generally 70-80%. The key point of the technical scheme is as follows: firstly, independently inoculating four microorganisms into potato residue (culture medium) in sequence for staged fermentation; secondly, when lactobacillus rhamnosus or lactobacillus casei is inoculated, saccharifying enzyme and liquefying enzyme are added in a matching manner, and the adding time of urea is prolonged; thirdly, appropriate culture conditions are adopted in each independent fermentation stage.

The optimization scheme of the method is as follows: the first stage fermentation condition is 30-37 ℃ closed culture for 12h, the second stage fermentation condition is 28-32 ℃ culture for 12h, and the third stage fermentation condition is 28-32 ℃ culture for 3-7 d. The materials are added and uniformly mixed with the potato dregs in each stage.

The invention also provides a method for utilizing the waste water slag from potato starch processing, which adopts the technical scheme as follows:

the method for utilizing the potato starch processing wastewater and the potato residues by utilizing the method for producing the protein feed by utilizing the potato residues is characterized in that the method comprises the following steps of: the inoculation of the lactobacillus rhamnosus or the lactobacillus casei uses a lactobacillus rhamnosus or lactobacillus casei seed solution prepared by culturing lactobacillus rhamnosus or lactobacillus casei by fresh wastewater generated in potato starch processing, the inoculation of the candida utilis uses a candida utilis seed solution prepared by culturing candida utilis by fresh wastewater generated in potato starch processing, and the inoculation of the saccharomyces cerevisiae uses a saccharomyces cerevisiae seed solution prepared by culturing saccharomyces cerevisiae by fresh wastewater generated in potato starch processing.

Earlier researches show that seed solutions of four bacteria including lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae can be efficiently prepared by taking potato starch processing wastewater as a culture medium, and the efficiency of the seed solutions is basically equal to or even superior to that of special synthetic culture media such as YPD and MRS culture media. Based on the method, the seed liquid is prepared from the potato starch processing wastewater and is used for producing the protein feed by fermenting the potato residues, so that the comprehensive utilization method of the two byproducts of the potato starch processing technology is provided, the feed production cost is reduced, and the efficiency of the integral resource treatment of the waste is improved.

The invention also provides an application of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae in producing protein feed by fermenting potato starch processing wastewater residues, and the technical scheme is as follows:

the application of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae in producing protein feed by fermenting potato starch processing wastewater residues is characterized in that: adding liquefying enzyme and saccharifying enzyme into fresh potato residues generated in potato starch processing, inoculating lactobacillus rhamnosus or lactobacillus casei, and fermenting for 12h in a closed manner at the temperature of 30-37 ℃; adding urea, inoculating candida utilis, and fermenting for 12 hours at the temperature of 28-32 ℃; finally inoculating saccharomyces cerevisiae, and fermenting for 3-7 days at 28-32 ℃.

The application belongs to the microbial fermentation engineering technology. Different from the prior application technology of the microorganism in producing the protein feed from the potato residue, the concept of compound fermentation by using mixed strains for utilizing a plurality of strains is adopted, sequential staged independent fermentation is adopted, and necessary auxiliary materials are added in different fermentation stages in a matching way, so that the effect of each stage and the smooth connection of an up-flow process and a down-flow process are ensured.

Compared with the prior art, the invention has the beneficial effects that: (1) the method for producing protein feed for animal feeding by using potato residue fermentation is characterized by that it uses fresh potato residue as raw material, selects three bacterial strains and makes them implement staged independent fermentation, and adds the necessary auxiliary agents, and adopts the optimum culture condition to ensure that the true protein content of fermented product is high, and meets the protein feed standard. The method is simple to operate, common in raw materials, easy to obtain and store four selected strains, and economical and efficient in potato residue resource utilization. Meanwhile, the four strains are industrial microorganisms of the Food and Drug Administration (FDA) certified GRAS (Generally Recognized as safe), and the biological safety of the obtained feed is guaranteed. (2) Provides a comprehensive utilization method of waste water and waste residue (potato residue) generated in potato starch processing. The comprehensive utilization method firstly breaks through the conception that the treatment of the waste water in the prior art only aims at reaching the discharge standard, improves the waste water to the resource utilization level, and skillfully combines the resource utilization with the resource utilization of the potato dregs into a whole. The method also breaks through the idea of classifying and treating the waste water and the waste residues (potato residues) in the prior art, not only reduces the production cost of the protein feed, but also provides an integral technical scheme for solving the problem of environmental pollution in potato starch processing by comprehensive utilization. The whole comprehensive utilization method can be directly connected with the existing potato starch processing technology, and the processing technology level of the potato starch is improved. And the whole comprehensive utilization method has economic cost and simple and easily-controlled operation conditions, and is particularly suitable for popularization and application in rural areas for processing potato starch. (3) Provides an application method of lactobacillus rhamnosus, lactobacillus casei, candida utilis and saccharomyces cerevisiae in producing protein feed by fermenting potato starch processing wastewater residues.

Detailed Description

Preferred embodiments of the present invention will be further described with reference to the following examples.

1. Material

Potato starch processing waste residue (potato residue): obtained from Tong Yu agriculture and potato industry Co Ltd in Mian Yang of Sichuan province. The processing technology of the potato starch comprises the steps of crushing sweet potatoes, adding water to prepare potato pulp, sieving by a 120-mesh sieve to obtain discarded waste residues (potato residues), and draining for later use; the powder slurry is sieved by a 200-mesh screen, and the waste residue (potato residue) is removed on the screen. In the experiment, the potato residues generated in the two processes are combined, and the water is drained for later use. The water content of the potato residue is measured to be 75.7 percent, the true protein content is measured to be 2.47 percent (DW), and the starch content is measured to be 53.2 percent (DW).

Potato starch processing wastewater: in the processing technology of the potato starch, after being filtered by a 200-mesh screen, the powder slurry is screened and discharged into a sedimentation tank for natural sedimentation overnight, the lower layer is the extracted starch, and the supernatant is the discarded wastewater. Collected for later use in this implementation.

Strain:

lactobacillus rhamnosus: lactobacillus rhamnosus CICC 20258, purchased from China center for Industrial culture Collection of microorganisms.

Lactobacillus casei: lactobacillus casei CICC 23184, purchased from China center for Industrial culture Collection of microorganisms.

Candida utilis: candida utilis CICC 1314 purchased from China center for culture collection and management of industrial microorganisms.

And (3) saccharomyces cerevisiae: saccharomyces cerevisiae CCTCC M206111, deposited in the China center for type culture Collection.

Saccharifying enzyme: suhong GA II: standard enzyme activity 500AGU ml from Novoxin^-1. AGU is a proprietary unit of Novoxil liquefying enzyme, and 1AGU refers to the amount of enzyme required to hydrolyze 1 microgram of maltose per minute at 25 ℃, pH 4.3, and a reaction time of 30 minutes.

Liquefying enzyme: liquozyme Supra, available from Novoxin, is a thermostable alpha-amylase with a standard enzyme activity of 90KNU/g (KNU is a proprietary unit of Novoxin liquefying enzyme). The definition of 1KNU is the amount of enzyme that hydrolyses 5.26g of starch per hour at 37 ℃ and pH 5.6.

Urea: purchased from a chemical reagent factory of Synechocystis.

MRS culture medium: taking 10.0g of casein peptone, 10.0g of beef extract, 5.0g of yeast powder, 20.0g of glucose, 5.0g of sodium acetate, 2.0g of diammonium citrate, 801.0 g of Tween and K₂HPO₄ 2.0g，MgSO₄.7H₂O 0.2g，MnSO₄.H₂O 0.05g，CaCO₃20.0g, 1.0L of distilled water, and pH adjusted to 6.8. Sterilizing at 115 deg.C for 20min, and cooling.

YPD medium: preparing YPD culture medium with glucose 20.0g, peptone 20.0g, yeast powder 10.0g, and distilled water 1.0L, sterilizing at 115 deg.C for 20min, and cooling.

2. Detection method and results

And (3) counting microorganisms: the blood cell counting method adopts a 16-lattice-by-25-lattice blood cell counting plate, and adopts the following calculation formula: cell count/ml-100 cells/100 × 400 × 10000 × dilution factor.

Detection of true protein: trichloroacetic acid is combined with a Kjeldahl method, a sample is firstly pickled for 3 times by 10 percent trichloroacetic acid, water-soluble nitrogen-containing substances such as urea and the like are leached out, the filtration is carried out, filter residues are dried, and then protein is measured by adopting a half-micro Kjeldahl method according to GB/T5009.5-2010.