CN101897381B - Method for producing microbial feed additive by utilizing potato distiller's grains - Google Patents

Abstract

The invention relates to a method for producing a microbial feed additive by using potato vinasse. The method comprises the following steps of: preparing a vinasse culture medium; mixing bacillus subtilis, trichoderma koningii and bacterial suspension liquid of trichoderma viride spores or mouldy bran, and adding into the vinasse culture medium for culturing and fermenting; and inoculating candida tropicalis or candida utilis seed liquid in the well-fermented culture medium for continuously culturing and fermenting; and finally drying to obtain the microbial feed additive. The invention has the advantages of low energy consumption, low cost, short production period, high production efficiency and recycled resources, reduces the emission of pollutants and is beneficial to building an environment-friendly and resource-saving society; and the produced additive has wide application range, can be added into animal feeds and is a high-quality and highly-efficient microbial fermentation product.

Description

Method for producing microbial feed additive by utilizing potato distiller's grains

technical field

The invention relates to a production method of a microbial feed additive, in particular to a method for producing a microbial feed additive by using the by-product potato distiller's grains after brewing alcohol in a brewery.

Background technique

With the increasing scale of ethanol production in the world, people are facing another problem: how to efficiently utilize alcohol waste residue and waste liquid to maximize the profit of the alcohol industry, reduce waste emissions, reduce environmental pollution, and achieve energy saving, emission reduction, The dual purpose of comprehensive utilization. Alcohol waste liquid is the main by-product of the winery, its volume is about 10 times the alcohol production. All countries in the world are trying to solve the problem of distiller's grain utilization. Potato distiller's grains mainly include potato distiller's grains, cassava distiller's grains, and sweet potato distiller's grains. The protein content of three potato distiller's grains is equivalent to that of barley, corn, and wheat distiller's grains, and the content of cellulose, total sugar and dry matter is higher than that of grain distiller's grains. This shows that the available range of potato distiller's grains is still very wide.

At present, there are many ways to use distiller's grains to produce feed: one is to use fresh distiller's grains as feed directly, but fresh distiller's grains tend to turn sour and are suitable for the growth of mold, so they cannot be stored for a long time. This means that the time for animals to feed distiller's grains is relatively short, and the water content of distiller's grains is high, and the transportation cost is high. Only users who live near the winery can use raw distiller's grains to feed livestock is cost-effective. This makes the method of directly utilizing distiller's grains as feed have limitations. The second is to concentrate and dry distiller's grains to prepare DDGS (Distiller's Dried Grains with Solubles, DDGS). Concentrated and dried starch-based distillers grains are generally accepted as feed in the United States and Canada. The solid phase is separated from the liquid using a sieve, centrifuge or extrusion device, the solid is dried (Distiller's Dried Grain, DDG), while the liquid is concentrated to contain 30-40% suspended solids (Distiller's Dried Solubles, DDS). In some distilleries the concentrate is dried and mixed with previously separated solids before drying (Distiller's Dried Grains with Solubles, DDGS). Both are characterized by high energy demands and this method is only suitable for very large wineries. Potato distiller’s grains are different from corn grains and rice grains in that they have deep color, high viscosity, and many fibers, and it is difficult to separate solids and liquids in the early stage. Therefore, the preparation of DDGS from potato distillers grains by means of concentration and drying will cost a lot due to equipment investment and drying concentration. The operating costs are so high that they are not economically viable. The third is to inoculate microorganisms to prepare fermented feed. Previous studies generally inoculate single strains such as lactic acid bacteria or yeast for fermentation to prepare lactic acid feed or single-cell protein feed. The feed obtained in this way can be stored for several months, prolonging the storage period or Improve its protein content, but the crude fiber content of distiller's grains is high, which limits the range of utilization of distiller's grains as feed, especially in non-ruminant farms.

Contents of the invention

The technical problem to be solved by the present invention is to provide a method for producing microbial feed additives with distiller's grains of potatoes with low energy consumption, wide application range and good quality in view of the above-mentioned deficiencies in the prior art.

In order to solve the above-mentioned technical problems, the technical solution adopted in the present invention is: a method for producing microbial feed additives using potato distiller's grains, the method comprising the following steps:

a, obtain fresh potato distiller's grains, bran and soybean meal, pulverize the bran and soybean meal respectively through a 40-mesh sieve; press the fresh potato distiller's grains: bran: soybean meal in a ratio of 5:3:2 by weight , bran and soybean meal are mixed, and 1.0-1.5% of (NH ₄ ) ₂ SO ₄ and 0.2-0.4% of KH ₂ PO ₄ are added to the mixture to make distiller's grain medium;

B, prepare Bacillus subtilis suspension, Trichoderma corning suspension, and Trichoderma viride spore suspension; Bacillus subtilis suspension, Trichoderma corning suspension, and Trichoderma viride spore suspension are by volume ratio 1: Mix at a ratio of 1-2:1-2, add the mixture to the above-mentioned distiller's grain medium at a ratio of 100 grams of distiller's grain medium plus 10-30 ml of mixed bacterial suspension, and culture and ferment at 28-30°C for 24-72 hours;

c. Prepare Candida tropicalis or Candida utilis seed liquid, inoculate Candida tropicalis or Candida utilis seed liquid in the above-mentioned fermented medium, and the inoculum amount is the inoculation of the medium fermented per 100 grams 10ml of Candida tropicalis or Candida utilis seed liquid, then cultured and fermented at a constant temperature of 28°C for 100-120h; dried at 50-55°C.

The b step of above-mentioned scheme also can adopt the method for preparing Bacillus subtilis gluten bent rice, Trichoderma koningen spore bran yeast, and Trichoderma virides spore bran; The koji is mixed according to the ratio of 1:1-2:1-2 by weight, and the mixed bran koji is added to the above-mentioned distiller's grain medium, and the weight is 10-30% of the distiller's grain medium, and it is cultivated and fermented at 28-30°C for 24 hours. -72h.

The fresh potato distiller's grains are obtained from a winery, which is a by-product of alcohol brewing in a winery, and can be any of potato distiller's grains, cassava distiller's grains, or sweet potato distiller's grains. The number of bacteria in the seed solution of Candida tropicalis or Candida utilis is 1.0-3.0×10 ⁶ /mL.

The above-mentioned Bacillus subtilis, Trichoderma konningen, Trichoderma viride, Candida tropicalis, and Candida utilis are known commonly used microorganisms, all of which can be purchased from the China Industrial Microbiology Collection Center market.

The Bacillus subtilis, Trichoderma konningen, and Trichoderma viride bran koji can be prepared by known methods: test tube strain → triangular flask seed koji → curtain seed koji → mechanical ventilation seed koji, the cellulase activity should reach 9000U/g bran song. For the specific process of making bran koji, please refer to "Food Microbiology" edited by He Guoqing, 2002.8 first edition: pages 251-265.

The preparation method of the seed solution of Candida tropicalis and Candida utilis is as follows (the preparation process of the two bacteria is the same): inoculate the slant strains of the bacteria stored in the refrigerator at 4°C into the fresh slant medium by streaking, 28 Cultivate to the logarithmic growth phase at ℃, pour a certain amount of sterile physiological saline, and wash off the bacterial lawn, so that the number of bacteria is 1.0-3.0×10 ⁶ /mL.

The production of Bacillus subtilis, Trichoderma koningeni, and Trichoderma viride suspensions can refer to the production of Candida tropicalis seed liquid above. The above-mentioned manufacturing methods are all known techniques in the art, and will not be repeated here.

A feed containing the microbial feed additive produced by the above-mentioned method, the feed additive is added in an amount of 1.0-2.0 kg per ton of feed.

Illustrate the application of the additive that the inventive method makes with concrete example below in animal feed:

The impact of feed additive of the present invention on piglet production performance:

A total of 90 weaned piglets aged 28 days were selected, randomly divided into 3 groups, and each group had 3 replicates. They were fed with 3 different rations, and had free access to food and water. The 1st group is a blank control group, and the 2nd and 3rd groups are respectively adding 0.1%, 0.2% additive of the present invention in the basal diet (0.1%, 0.2% are percentage by weight, that is, the per ton of feed mentioned above Add 1-2kg additive of the present invention).

The basic diet is composed of: corn 59.6%, soybean meal 20.0%, fish meal 5.0%, whey powder 10.0%, palm oil 2.0%, stone powder 0.8%, calcium hydrogen phosphate 1.0%, salt 0.25%, enzyme preparation 0.2%, anti-mold Agent 0.15%, and premix 1.0%.

The main nutritional levels are: digestible energy 13.99MJ/kg, crude protein 18.4%, calcium 0.94%, phosphorus 0.68%, salt 0.36%, lysine 1.17%. The diet was designed according to the nutritional needs of NRC10-20kg piglets, and the test period was one month. The results are shown in Table 1 below:

Table 1 Effect of microbial feed additive of the present invention on growth performance of weaned piglets

It can be seen from the above table that the average final weight of piglets in the 2nd and 3rd groups increased by 7% and 17% respectively compared with the control group. Analysis of variance showed that the ADG (average daily gain) of each group was significantly different, and the ADG of the 2nd and 3rd groups increased by 12% and 29% respectively compared with the control group, and adding 0.2% microbial feed additives could significantly improve the daily gain of piglets; Group ADFI (daily feed intake), F/G (feed-to-weight ratio) difference is not significant, but as can be seen from the table, the F/G of the 2nd and 3rd groups is lower than the control group by 12%, 15% respectively; The diarrhea rate of piglets in the 2nd and 3rd groups was significantly lower than that of the control group, which was 70% and 48% lower than the control group respectively, while the difference between the 2nd and 3rd groups was not significant.

The present invention has the following advantages:

1. According to the high cellulose content of potato distiller's grains, first use the cellulose degrading bacteria made from the suspension of Bacillus subtilis, Trichoderma korningii, and Trichoderma viride spores or the mixture of glutinous rice bran to degrade the cellulose, and then Inoculate yeast for fermentation process, which solves the defects of high cellulose content, poor palatability and narrow application range of distiller's grain feed;

2. According to the characteristics of high viscosity of fresh distiller's grains, the present invention changes cellulase into bran koji, which not only saves the cost brought by the use of a large amount of enzyme preparations, but also reduces the viscosity of distiller's grains, accelerates the cellulose degradation process, and shortens the production cycle. Increase productivity;

3. According to the high water content of fresh distiller's grains, adding a certain amount of bran and soybean meal not only increases the nutrients necessary for the growth of microorganisms, but also saves energy without drying the distiller's grains; and does not need to add additional water, saving water is a one-shot Three good ways;

4. Distiller's grain medium is inoculated with yeast for feed, which increases the protein content of the feed and the number of beneficial microorganisms. It also contains active components of Bacillus and yeast physiological metabolism. Adding it to animal feed can promote the absorption of nutrients and increase body weight and improved feed conversion efficiency;

5. The additive of the present invention has a wide application range and can be added to the feed of pigs, cattle, chickens, ducks, fish and other animals, and is a high-quality and efficient microbial fermentation product for feed. Moreover, the energy consumption is low; the cost is low, the production cycle is short, and the production efficiency is high; it can achieve resource recycling and reduce pollutant discharge, which is conducive to building an environment-friendly and resource-saving society.

Detailed ways:

The present invention will be further described below in conjunction with embodiment. These examples should not be used to interpret the limitations of the protection scope of the present invention.

Obtain fresh potatoes, cassava, and sweet potato distiller's grains from the brewery; respectively pulverize commercially available bran and soybean meal, and pass through a 40-mesh sieve; liquid and bran yeast, and Trichoderma viride suspension and gluten gluten; and prepared Candida tropicalis or Candida utilis seed liquid according to the prior art; to be used in the following examples.

Example 1:

The weight ratio of fresh potato distiller's grains: bran: soybean meal is 5:3:2, fresh potato distiller's grains, bran and soybean meal are mixed, and 1.0% of (NH ₄ ) is added to the total weight of fresh potato distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.2% KH ₂ PO ₄ to make distiller's grain medium, the prepared Bacillus subtilis suspension, Trichoderma korningii suspension, and Trichoderma viride suspension were prepared in a volume ratio of 1:1:1 After mixing, add in the above-mentioned culture medium, add 20ml per 100 grams of distiller's grain culture medium, after culturing and fermenting for 72h at 28°C, inoculate 10ml of activated Candida tropicalis ( Inoculate Candida tropicalis at a ratio of 1.6×10 ⁶ cells/mL), continue to culture at 28°C for 120 hours, and then dry it. It was determined that the protein content of the product increased by 42.8%, the cellulose content decreased by 8.3%, the number of bacillus was 2.1×10 ⁹ /mL, and the number of yeast was 1.7×10 ⁷ /mL.

Example 2:

According to the weight ratio of fresh cassava distiller's grains: bran: soybean meal is 5:3:2, fresh cassava distiller's grains, bran and soybean meal are mixed, and 1.5% (NH ₄ ) is added to the total weight of fresh cassava distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.2% KH ₂ PO ₄ to make distiller's grain medium, the prepared Bacillus subtilis suspension, Trichoderma korningen suspension, and Trichoderma viride suspension in a volume ratio of 1:1:1 After mixing, add in the above-mentioned culture medium, add 20ml per 100 grams of distiller's grain culture medium, after culturing and fermenting for 48h at 29°C, inoculate 10ml of activated Candida tropicalis ( Inoculate Candida tropicalis at a ratio of 3.0×10 ⁶ cells/mL), continue to culture at 28°C for 120 hours, and then dry it. It was determined that the protein content of the product increased by 37.8%, the cellulose content decreased by 7.6%, the number of bacillus was 1.9× ¹⁰⁸ /mL, and the number of yeast was 2.7× ¹⁰⁷ /mL.

Example 3:

According to the weight ratio of fresh sweet potato distiller's grains: bran: soybean meal is 5:3:2, fresh sweet potato distiller's grains, bran and soybean meal are mixed, and 1.2% (NH ₄ ) is added to the total weight of fresh sweet potato distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.2% KH ₂ PO ₄ to make a distiller's grain medium, and mix the prepared Bacillus subtilis bran koji, Trichoderma korningina bran koji, and Trichoderma viride koji in a weight ratio of 1:1:1 Then add it to the above-mentioned medium, add 20 grams per 100 grams of distiller's grains medium, culture and ferment at 28 ° C for 24 hours, then inoculate 10 ml of activated Candida utilis in the fermented medium per 100 grams of medium (the number of bacteria is 1.0×10 ⁶ cells/mL) to inoculate Candida utilis, continue to culture at 28°C for 120h, and then dry it. It was determined that the protein content of the product increased by 38.4%, the cellulose content decreased by 7.9%, the number of bacillus was 2.0×10 ⁹ /mL, and the number of yeast was 1.2×10 ⁷ /mL.

Example 4:

The weight ratio of fresh sweet potato distiller's grains: bran: soybean meal is 5:3:2, fresh sweet potato distiller's grains, bran and soybean meal are mixed, and 1.5% (NH ₄ ) is added to the total weight of fresh sweet potato distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.3% KH ₂ PO ₄ to make distiller's grains medium, the prepared Bacillus subtilis suspension, Trichoderma korningii suspension, and Trichoderma viride suspension in a volume ratio of 1:1:1 After mixing, add in the above-mentioned culture medium, add 30ml per 100 grams of distiller's grain culture medium, after culturing and fermenting at 28°C for 48h, inoculate 10ml of activated Candida tropicalis ( Inoculate Candida tropicalis at a ratio of 1.6×10 ⁶ cells/mL), continue culturing at 28°C for 100 h, and then dry it. It was determined that the protein content of the product increased by 35.4%, the cellulose content decreased by 9.2%, the number of bacillus was 1.5× ¹⁰⁸ /mL, and the number of yeast was 1.3× ¹⁰⁷ /mL.

Example 5:

According to the weight ratio of fresh cassava distiller's grains: bran: soybean meal is 5:3:2, fresh cassava distiller's grains, bran and soybean meal are mixed, and 1.0% (NH ₄ ) is added to the total weight of fresh cassava distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.4% KH ₂ PO ₄ to make a distiller's grain medium, and mix the prepared Bacillus subtilis bran koji, Trichoderma korningina bran koji, and Trichoderma viride koji bran in a weight ratio of 1:2:1 Then add it to the above medium, add 30 grams per 100 grams of distiller's grains medium, culture and ferment at 30 ° C for 24 hours, then inoculate 10 ml of activated Candida utilis into the fermented medium per 100 grams of medium (the number of bacteria is 3.0×10 ⁶ cells/mL) to inoculate Candida utilis, continue to culture at 28°C for 100h, and then dry it. It was determined that the protein content of the product increased by 36.7%, the cellulose content decreased by 8.1%, the number of bacillus was 3.1× ¹⁰⁸ /mL, and the number of yeast was 2.4× ¹⁰⁷ /mL.

Embodiment 6:

The ratio by weight of fresh cassava distiller's grains: bran: soybean meal is 5:3:2, fresh cassava distiller's grains, bran and soybean meal are mixed, and 1.2% (NH ₄ ) is added to the total weight of fresh cassava distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.3% KH ₂ PO ₄ to make distiller's grain medium, the prepared Bacillus subtilis suspension, Trichoderma korningii suspension, and Trichoderma viride suspension were prepared in a volume ratio of 1:1:2 After mixing, add in the above-mentioned culture medium, add 10ml per 100 grams of distiller's grain culture medium, after culturing and fermenting for 72 hours at 28°C, inoculate 10ml of activated Candida tropicalis ( Inoculate Candida tropicalis at a ratio of 1.6×10 ⁶ cells/mL), continue to culture at 28°C for 120 hours, and then dry it. It was determined that the protein content of the product increased by 37.6%, the cellulose content decreased by 8.0%, the number of bacillus was 2.7× ¹⁰⁸ /mL, and the number of yeast was 2.3× ¹⁰⁷ /mL.

Embodiment 7:

According to the weight ratio of fresh potato distiller's grains: bran: soybean meal is 5:3:2, fresh potato distiller's grains, bran and soybean meal are mixed, and 1.2% (NH ₄ ) is added to the total weight of fresh potato distiller's grains, bran and soybean meal ₂ SO ₄ , and 0.4% KH ₂ PO ₄ to make a distiller's grain medium, and mix the prepared Bacillus subtilis bran koji, Trichoderma korningina bran koji, and Trichoderma viride koji bran in a weight ratio of 1:2:2 Then add it to the above medium, add 20 grams per 100 grams of distiller's grains medium, culture and ferment at 29 ° C for 48 hours, then inoculate 10 ml of activated Candida utilis per 100 grams of medium in the fermented medium (The number of bacteria is 1.0×10 ⁶ cells/mL) to inoculate Candida utilis, continue culturing at 28°C for 110 hours, and then dry it. It was determined that the protein content of the product increased by 38.4%, the cellulose content decreased by 10.2%, the number of bacillus was 2.0× ¹⁰⁸ /mL, and the number of yeast was 1.2× ¹⁰⁷ /mL.

Claims (7)

1. A method utilizing potato distiller's grains to produce microbial feed additives, characterized in that: the method may further comprise the steps: a, obtain fresh potato distiller's grains, bran and soybean meal, pulverize the bran and soybean meal respectively through a 40-mesh sieve; press the fresh potato distiller's grains: bran: soybean meal in a ratio of 5:3:2 by weight , bran and soybean meal are mixed, and 1.0-1.5% of (NH ₄ ) ₂ SO ₄ and 0.2-0.4% of KH ₂ PO ₄ are added to the mixture to make distiller's grain medium; B, prepare Bacillus subtilis suspension, Trichoderma corning suspension, and Trichoderma viride spore suspension; Bacillus subtilis suspension, Trichoderma corning suspension, and Trichoderma viride spore suspension are by volume ratio 1: Mix at a ratio of 1-2:1-2, add the mixture to the above-mentioned distiller's grain medium at a ratio of 100 grams of distiller's grain medium plus 10-30 ml of mixed bacterial suspension, and culture and ferment at 28-30°C for 24-72 hours; c. Prepare Candida tropicalis or Candida utilis seed liquid, inoculate Candida tropicalis or Candida utilis seed liquid in the above-mentioned fermented medium, and the inoculum amount is the inoculation of the medium fermented per 100 grams 10ml of Candida tropicalis or Candida utilis seed liquid, then cultured and fermented at a constant temperature of 28°C for 100-120h; dried at 50-55°C. 2. The method of utilizing potato distillers grains to produce microbial feed additives as claimed in claim 1, characterized in that: said fresh potato distillers grains are by-products after brewing alcohol in distilleries, and are potato distillers grains, cassava distillers grains, or sweet potato distillers grains A type of distiller's grains. 3. The method for producing microbial feed additives using potato distiller's grains as claimed in claim 1, characterized in that: the number of bacteria in the seed liquid of Candida tropicalis or Candida utilis is 1.0-3.0× ¹⁰⁶ / mL. 4. A method utilizing potato distiller's grains to produce microbial feed additives, characterized in that: the method may further comprise the steps: a, obtain fresh potato distiller's grains, bran and soybean meal, pulverize the bran and soybean meal respectively through a 40-mesh sieve; press the fresh potato distiller's grains: bran: soybean meal in a ratio of 5:3:2 by weight , bran and soybean meal are mixed, and 1.0-1.5% of (NH ₄ ) ₂ SO ₄ and 0.2-0.4% of KH ₂ PO ₄ are added to the mixture to make distiller's grain medium; B, prepare Bacillus subtilis bran yeast, Trichoderma konningen spore bran yeast, and Trichoderma viride spore bran yeast; with Bacillus subtilis bran yeast, Trichoderma konningen spore bran yeast, and Trichoderma viride spore bran yeast in a weight ratio of 1: 1- Mixing at a ratio of 2:1-2, adding the mixed bran koji to the above-mentioned distiller's grains medium, the added weight is 10-30% of the weight of the distiller's grains medium, and cultivating and fermenting at 28-30°C for 24-72h; c. Prepare Candida tropicalis or Candida utilis seed liquid, inoculate Candida tropicalis or Candida utilis seed liquid in the above-mentioned fermented medium, and the inoculum amount is the inoculation of the medium fermented per 100 grams 10ml of Candida tropicalis or Candida utilis seed liquid, then cultured and fermented at a constant temperature of 28°C for 100-120h; dried at 50-55°C. 5. The method for producing microbial feed additives using potato distiller's grains as claimed in claim 4, characterized in that: said potato distiller's grains are by-products after brewing alcohol in distilleries, and are potato distiller's grains, cassava distiller's grains, or sweet potato distiller's grains One of. 6. The method for producing microbial feed additives using potato distiller's grains as claimed in claim 4, characterized in that: the number of bacteria in the seed liquid of Candida tropicalis or Candida utilis is 1.0-3.0× ¹⁰⁶ / mL. 7. A feed containing a microbial feed additive, characterized in that: the feed is added with the feed additive prepared by the method according to any one of the above claims, and the amount of the feed additive is 1.0-2.0 per ton of feed kg.