CN104388317A - A kind of organic chlorella production method rich in trace elements - Google Patents

Abstract

The invention discloses a method for producing organic chlorella rich in trace elements, which comprises the steps of preparing a microbial anaerobic fermentation medium, anaerobic fermentation of liquid organic fertilizer and producing organic chlorella. The invention is suitable for experimental production above the pilot level and large-scale chlorella production. The invention realizes the production of liquid organic fertilizer of chlorella through the anaerobic fermentation method, and the enrichment and organicization of trace elements are realized in the link of configuring the chlorella cultivation medium. The liquid fertilizer rich in organic trace elements is used to cultivate chlorella, so that an organic chlorella product enriched with various trace elements can be obtained. The various trace elements in the chlorella product have been organicized and are more easily absorbed by the human body. The invention uses biomass waste as source material, realizes the resource utilization and high value of waste, and the organicization of the chlorella production process brings both environmental and economic effects.

Description

A kind of organic chlorella production method rich in trace elements

technical field

The invention relates to a production method of organic chlorella rich in trace elements. Specifically, the liquid organic fertilizer rich in trace elements is used to cultivate chlorella rich in iron, zinc, iodine, chromium, selenium and other trace elements, which belongs to the field of bioengineering. the

Background technique

Iron, zinc, copper, manganese, chromium, selenium, molybdenum, cobalt, fluorine and other elements only account for less than 0.1% of the total weight in the human body. These elements are called trace elements of the human body. Trace elements are closely related to the health of the human body. vital role in life. According to research, so far, 18 kinds of essential trace elements have been confirmed to be related to human health and life, namely iron, copper, zinc, cobalt, manganese, chromium, selenium, iodine, nickel, fluorine, molybdenum, vanadium, Tin, silicon, strontium, boron, rubidium, arsenic, etc., although their content in the human body is very small, they are indispensable substances to maintain some decisive metabolism in the human body, such as iron is the prosthetic group of heme enzyme, iodine Copper is the cofactor of cytochrome oxidase, manganese is the cofactor of arginase, zinc is the cofactor of deoxygenase, DNA polymerase, carbonic anhydrase, selenium is the cofactor of glutathione cofactor of oxidase, etc., cobalt is a component of vitamin B12, vanadium is a cofactor of nitrate reductase, molybdenum is a cofactor of xanthine oxidase, and nickel is a cofactor of urease. Excessive, insufficient, unbalanced or deficient intake of trace elements will cause abnormalities in human physiology or diseases to varying degrees. At present, it is relatively clear that about 30% of diseases are directly caused by the lack or imbalance of trace elements. At the same time, studies have found that trace elements still play a very important role in disease resistance, cancer prevention, and longevity.

Trace elements mainly exist in soil and rocks in nature, and exist in the form of inorganic substances. The trace elements needed by the human body need to be ingested through diet, but in actual life, the absorption efficiency of the inorganic form of trace elements by the human body is not high. Most people often do not get enough trace elements through diet and need to make additional compensation.

Chlorella is a natural food with balanced nutrition. It has multi-functional and health-care effects. It contains a variety of nutrients necessary for human growth, health and longevity. Chlorella has high protein content and balanced amino acid content. It meets the nutritional needs of the human body, and its protein peptide has a variety of biological activities, which can activate human cells, repair damaged genes, and promote the growth of organ tissues; Free radicals, anti-aging, improving heart and lung functions and other functions, chlorella polysaccharides and a variety of trace elements, have anti-fatigue, anti-atherosclerosis, prevent cardiovascular disease and diabetes and other effects; chlorella unique CGF active factor has the functions of regulating immune function, anti-tumor, inducing interferon, and enhancing cell phagocytosis; the natural high-concentration chlorophyll and plant cellulose in Chlorella can eliminate chemical toxins and heavy metals in the body, protect the liver and kidney from Toxicity, cancer prevention and other effects.

During the growth process of Chlorella, a remarkable biological feature is that it can enrich the trace element metal ions in the culture medium, and has the function of converting the inorganic trace elements in the environment into organic trace elements, which is more beneficial to the human body Absorption, but in actual production, chlorella culture mostly adopts the method of photoautotrophy, directly adding inorganic trace elements to the medium, it is difficult to control the concentration of trace elements in the algae cells, and a considerable part is only adsorbed on the cell surface, It is still in an inorganic state and is not easily absorbed by the human body; in addition, there may still be a large amount of trace element residues in the water body of the culture medium, and the discharge may easily cause environmental pollution.

The invention provides a production method of organic chlorella rich in trace elements, utilizing the biological characteristics that chlorella can carry out heterotrophic culture with organic matter, and producing enriched organic trace elements by the method of microbial cell fermentation The medium, which is further formulated and used for chlorella cultivation, can produce chlorella rich in various trace elements. the

Contents of the invention

The purpose of the present invention is to provide a method for producing chlorella rich in trace elements for the deficiencies of the prior art, which can be used to cultivate and produce chlorella rich in various trace elements, and more accurately Control the content of trace elements in the algae body, and the degree of organicization of the trace elements in the algae body is high, which is more conducive to human body absorption, improves the quality of chlorella products, and realizes the diversification, organicization and further high value of chlorella products . In addition, the method utilizes industrial and agricultural waste biomass as the main raw material, realizes the recycling of waste, and has obvious environmental and social benefits

In order to achieve the above object, the present invention is achieved through the following technical solutions.

A method for producing organic chlorella rich in trace elements, comprising the following process steps, specifically:

a. Preparation of microbial anaerobic fermentation medium: use waste biomass as the main raw material, make ingredients according to its nutrient components, supplement nitrogen, phosphorus, potassium and other large elements according to demand to adjust the balance of various elements in the medium, and add zinc, chromium ( Cr ³⁺ ), selenium, iodine and other inorganic salts are used as trace element ingredients; the raw materials are crushed and mixed, then steam sterilized, and sterile water is added to prepare an anaerobic fermentation medium;

b. Anaerobic fermentation of liquid organic fertilizer: add cellulolytic bacteria and proteolytic bacteria strains to the culture medium prepared in step a, and ferment under anaerobic conditions to obtain Fermentation liquid of amino acids, protein peptides, bacteria and other organic matter, the fermentation liquid is also rich in zinc, chromium (Cr ³⁺ ), selenium, iodine and other trace elements;

c. Production of organic chlorella: sterilize the fermented liquid rich in trace elements obtained in step b, after deployment, add it into the chlorella culture container for chlorella cultivation; After harvesting and drying, organic chlorella products enriched with various trace elements are obtained, which can be used for further processing and development.

Among them, the waste biomass mentioned in step a refers to various wastes and by-products produced in industrial and agricultural production, containing protein, polysaccharides (mainly starch and cellulose) and other biomass resources that can be fermented by microorganisms, such as fermentation Wastewater, brewing wastewater, distiller's grains, cassava residue, bagasse, soybean meal, peanut meal, rice bran, wheat bran, agricultural straw, etc.

Among them, the nutritional components described in step a mainly refer to substances such as proteins and polysaccharides in waste biomass that can be utilized by microorganisms; equal component content, proportion raw materials according to a certain proportion, and correspondingly add fertilizers such as nitrogen, phosphorus, potassium, etc., to reach the nutrient and element proportion suitable for microbial fermentation growth; the addition of trace elements described in step a refers to According to the needs of the target chlorella products, correspondingly add zinc, chromium (Cr ³⁺ ), selenium, iodine and other inorganic salts of trace elements.

Wherein, the liquid organic fertilizer anaerobic fermentation described in step b refers to the process of cultivating cellulolytic bacteria, amylolytic bacteria, lipolytic bacteria and proteolytic bacteria in the culture medium prepared in step a under anaerobic conditions The principle is to use cellulolytic bacteria, amylolytic bacteria and lipolytic bacteria to decompose cellulose, starch, fat and other substances in the medium during the growth and reproduction process in the medium to become other fungi or chlorella that can be directly used Monosaccharides, oligosaccharides, low-yuan organic acids, etc.; use proteolytic bacteria to decompose protein substances during the growth and reproduction process in the medium, such as bacteria, proteins and other substances into free amino acids that Chlorella can directly use , The process of small protein peptides.

Further, the cellulolytic bacteria described in step b refers to the ability to decompose cellulose in biomass into monosaccharides, oligosaccharides and formic acid, acetic acid, and propionic acid under anaerobic conditions through bacterial production and metabolism. , butyric acid and other low-carbohydrate strains, such as various strains of Clostridium sporogenes.

Further, the amylolytic bacteria described in step b refers to the bacteria that can decompose the starch in the biomass into monosaccharides, oligosaccharides and formic acid, acetic acid, propionic acid, butyric acid under anaerobic conditions through the production and metabolism of bacteria. Sour and other low-carbohydrate strains, such as some strains of Bacillus subtilis.

Further, the lipolytic bacteria described in step b refers to the bacteria that can decompose the fat in biomass into monosaccharides, oligosaccharides and formic acid, acetic acid, propionic acid, butyric acid under anaerobic conditions through bacterial body production and metabolism. Sour and other low-carbohydrate strains, such as Clostridium sporogenes and Bacillus subtilis.

Further, the proteolytic bacteria described in step b refers to bacteria that can decompose proteins in biomass into free amino acids and small protein peptides through bacterial production and metabolism under anaerobic conditions. Most anaerobic bacteria in the environment Oxygen bacteria have the function of decomposing protein.

Wherein, the chlorella cultivation described in step c refers to using the fermented liquid rich in monosaccharides, oligosaccharides, low-basic organic acids, free amino acids, small protein peptides and various trace elements obtained in step b as a culture medium, The process of growing chlorella in a culture container as a raw material for chlorella culture. Further, the chlorella culture container refers to a fermenter, a closed photobioreactor, an open pool, and a series combination of the above different containers. the

Wherein, the centrifugal harvesting described in step c refers to the process of using a disc centrifuge to centrifugally harvest the chlorella to obtain a concentrated chlorella slurry.

Wherein, the drying described in step c refers to the process of using a spray drying tower to dry the chlorella thick slurry to obtain the chlorella dry powder.

The beneficial effects of the present invention are: a method for producing organic chlorella rich in trace elements according to the present invention, which includes the preparation of microbial anaerobic fermentation medium, the anaerobic fermentation of liquid organic fertilizer and the production of organic chlorella step. Through the above process steps, organic chlorella products rich in various trace elements are produced and obtained. The present invention is suitable for experimental production and large-scale chlorella production above the pilot test level; the present invention realizes the production of liquid organic fertilizer for chlorella through the method of anaerobic fermentation, and is configured in the chlorella culture medium The link realizes the enrichment and organicization of trace elements. Using this liquid fertilizer rich in organic trace elements to cultivate chlorella, one can obtain organic chlorella products enriched with various trace elements. The chlorella products The trace elements in the milk have been organicized and are more easily absorbed by the body. The invention utilizes the biomass waste as the source material, realizes waste resource and high value, and organicizes the production process of chlorella, bringing double effects of environment and economy.

Detailed ways

The present invention will be described below in conjunction with specific embodiments, wherein the chlorella referred to in the present invention refers to various strains of Chlorella defined as Chlorella in taxonomy, and the equipment used in the present invention Both refer to pilot scale and above-scale equipment.

Example 1, brewer's grains were used as the main raw material to configure microbial anaerobic fermentation medium. Brewer's grains were taken from Dongguan Kingway Brewery. After analysis, the dry matter contained 24.7% of crude protein, 7.5% of crude fat, and 14.2% of crude fiber; Take 700 kg of brewer's grains with a water content of 45% and add them to a 3-ton stainless steel sterilization tank, add 50 kg of molasses, add 750 kg of water, stir and mix well, pass steam, heat and sterilize at 120°C for more than 0.5 hours, and cool down to room temperature , into a sterilized 5-ton plastic bucket.

Take 20 kg of urea, 15 kg of salt, 10 kg of potassium dihydrogen phosphate, 15 kg of magnesium sulfate heptahydrate, 2 kg of ferric chloride, 3 kg of zinc sulfate heptahydrate, 0.75 g of sodium selenite, 1 g of potassium iodate, nitric acid Chromium [Cr(NO ₃ ) ₃ ] 0.75 g was fully melted and sterilized by filtration, then added to a 5-ton plastic bucket, and sterile water was added until the total volume was 3.5 tons, and the microbial anaerobic fermentation medium was configured.

Embodiment two, the production of chlorella liquid organic fertilizer, in the microorganism anaerobic fermentation culture medium that embodiment one obtains, in the 5 tons of plastic vats that liquid filling capacity is 3.5 tons, by the addition amount of 10g per ton of culture medium, Add commercially available EM bacteria powder, which contains bifidobacteria, lactic acid bacteria, bacillus, photosynthetic bacteria, yeast, actinomycetes, acetic acid bacteria and other bacteria species, and the total number of bacteria per gram is ≥ 50 billion CFU. After the plastic bucket is covered and sealed, it is placed outdoors, and the anaerobic bacteria in the EM bacteria powder use natural temperature conditions to grow, and the anaerobic fermentation time is more than 7 days. Take the fermentation supernatant and sterilize it at 120°C for 0.5 hours to obtain concentrated liquid organic fertilizer, which is rich in trace elements such as iron, zinc, selenium, trivalent chromium and iodine. Due to the uncontrolled anaerobic fermentation conditions, the concentration of each nutrient element in different batches of organic fertilizers is variable, and the nitrogen element content needs to be measured as a reference index, which is used as the basis for configuring the chlorella organic medium after dilution in the next step.

Embodiment 3, Chlorella heterotrophic fermentation culture, this embodiment uses 10 tons fermenter tank as culture container, adds sterile heterotrophic fermented Chlorella liquid as seed liquid and aseptic concentrated liquid organic fertilizer, supplements Sterile water, to a reaction volume of 7 tons. Control the initial fermentation liquid, the dry weight density of Chlorella is 0.25g±0.05/L, the final concentration of nitrogen in the medium is 0.22-0.25gN/L, the parameters of fermentation culture: stirring rate 180-200rpm, dissolved oxygen control 50±5%, the fermentation temperature is 28±0.5℃, and the pH is controlled at 7±0.2. After 24 hours of fermentation, the algae liquid is harvested and concentrated by disc centrifuge at a centrifugal rate of 4500rpm or more. After repeated cleaning- After centrifugation for 3 times, the concentrated solution (with a dry weight content of about 10%) is dried in a spray drying tower, and the drying temperature is controlled at 180°C. After drying, the organic chlorella algae powder produced by fermentation is obtained. After testing, the dry weight content of chlorella in the fermentation broth is 0.82g/L, the protein content in the organically produced chlorella is 48.7%, the water content is <5%, the ash content is <7%, the zinc content is 5.2mg/g, and the trivalent The chromium content is 22 μg/g, the iron content is 2.5 mg/g, the selenium content is 19 μg/g, and the iodine content is 31 μg/g.

Embodiment four, utilizing the closed photobioreactor to cultivate Chlorella, the present embodiment takes the sterilized vertical bag photobioreactor as the culture container, the diameter of the vertical bag is 25cm, the height is 2.2 meters, and the height of the working fluid is 1.75 meters, the working volume is 85L, 12 stand-up bags as a group, the total working volume is 1 cubic meter. Add chlorella seed liquid and sterile concentrated liquid organic fertilizer, supplement sterile water, control the initial culture medium, the dry weight density of chlorella is 0.15g±0.05/L, and the final concentration of nitrogen in the medium is 0.12 ～0.15gN/L, inflate and stir, the aeration ratio is 0.05vvm, under natural light, fill in carbon dioxide gas to supplement the carbon source, and control the pH to 8±0.2, add concentrated liquid organic fertilizer once every morning and afternoon to cultivate The final concentration of nitrogen in the liquid is ~0.15gN/L, cultivated outdoors, and produced using natural light. During the measurement and cultivation period, the water temperature is 22-31°C, and the maximum light intensity is 55-65klux. After three days of cultivation, the algae liquid is harvested, concentrated by centrifugation in a disc centrifuge, and the centrifugation speed is above 4500rpm, and then washed and centrifuged repeatedly for 3 times. Finally, the concentrated solution (dry weight content is about 10%) is dried in a spray drying tower, and the drying temperature is controlled at 180°C. After drying, the organic chlorella algae powder is obtained. After testing, the dry weight content of chlorella in the fermentation broth is 1.25g/L, the protein content in the organically produced chlorella is 61.2%, the water content is <5%, the ash content is <7%, the zinc content is 6.4 mg/g, and the trivalent The chromium content is 26 μg/g, the iron content is 3.2 mg/g, the selenium content is 23 μg/g, and the iodine content is 37 μg/g.

Embodiment five, utilize fermentor and stand-up bag reactor to carry out chlorella culture in series, present embodiment uses 10 ton level fermenter as culture vessel, culture steps and parameters are the same as embodiment three, after fermenting for 24 hours, transfer to In the vertical bag photobioreactor, the cultivation steps and parameters are the same as those in Example 4. After 24 hours of cultivation (inoculation at 18:00 p.m., after one exposure to sunlight), the algae liquid is harvested and concentrated by centrifugation with a disc centrifuge at a centrifugation rate of Above 4500rpm, after repeated washing and centrifugation for 3 times, the concentrated solution (dry weight content is about 10%) is dried in a spray drying tower, and the drying temperature is controlled at 180°C. Organic chlorella algae powder can be obtained after drying. After testing, the dry weight content of chlorella in the fermentation broth is 1.02g/L, the protein content in the organically produced chlorella is >60.2%, the water content is <5%, the ash content is <7%, and the zinc content is 5.7 mg/g. The valent chromium content is 23 μg/g, the iron content is 2.9 mg/g, the selenium content is 21 μg/g, and the iodine content is 34 μg/g.

Embodiment six, carry out chlorella cultivation in the open pond, take the open circular pond of 1000 square meters as the culture container, add the water of 10cm depth, add the concentrated liquid organic fertilizer of chlorella seed liquid, add water to the cultivation depth is 20cm, Control the initial culture medium, the dry weight density of chlorella is 0.12g±0.02/L, the final concentration of nitrogen in the culture medium is , use a mixer truck to stir, the speed is every 72s, and add concentrated organic fertilizer once every morning Until the final concentration of nitrogen in the medium is 0.12-0.15gN/L. Cultivated outdoors with natural light for production. During the measurement and cultivation period, the water temperature is 24-32°C, and the maximum light intensity is 58-67klux. After four days of cultivation, the algae liquid is harvested, concentrated by centrifugation in a disc centrifuge, and the centrifugation speed is above 4500rpm, and then washed and centrifuged repeatedly for 3 times. Finally, the concentrated solution (dry weight content is about 10%) is dried in a spray drying tower, and the drying temperature is controlled at 180°C. After drying, the organic chlorella algae powder is obtained. After testing, the dry weight content of chlorella in the fermentation broth is 0.58g/L, the protein content in the organically produced chlorella is 59.7%, the water content is <5%, the ash content is <7%, the zinc content is 6.4 mg/g, and the trivalent The chromium content is 32 μg/g, the iron content is 3.5 mg/g, the selenium content is 27 μg/g, and the iodine content is 42 μg/g.

The above content is only a preferred embodiment of the present invention. For those of ordinary skill in the art, according to the idea of the present invention, there will be changes in the specific implementation and application range, such as the types of added trace elements and ingredients. The ratio of each raw material, the change of each parameter in the fermentation culture, the application concentration of organic fertilizer in the cultivation process, etc., the content of this specification should not be understood as a limitation of the present invention. the

Claims (4)

1. an organic chlorella production method for rich trace elements, is characterized in that: it comprises the following steps, and the first step is the preparation of microbiological anaerobic fermention medium; Second step is producing of fertilizer anaerobic fermented liquid; 3rd step is in culture vessel, using the cultivation raw material of fertilizer anaerobic fermented liquid as chlorella, adds chlorella seed liquor and carries out chlorella cultivation, carries out organic chlorella product that drying can obtain rich trace elements after results algae liquid.

2. organic chlorella production method of a kind of rich trace elements according to claim 1, it is characterized in that: the preparation of microbiological anaerobic fermention medium, utilize abandoned biomass for main raw material, prepare burden according to its nutrition composition, supplement the fertilizer such as nitrogen, phosphorus, potassium and regulate substratum various element balance, add zinc, chromium (Cr simultaneously ³⁺), the inorganic salt element such as selenium, iodine prepares burden as trace element; Pass into steam sterilizing after being pulverized by above-mentioned compound, add sterilized water and be deployed into anaerobically fermenting substratum; Described abandoned biomass, refer to the waste that all kinds of industrial and agricultural production produces and by product, the biomass of fermentable can be supplied, as fermentation waste water, brewing wastewater, vinasse, manioc waste, bagasse, dregs of beans, peanut meal, rice bran, wheat bran, agricultural stalk etc. containing protein, polysaccharide (being mainly starch and Mierocrystalline cellulose) etc.; Described nutrition composition, refers to the material such as protein and polysaccharide that can be utilized by microorganism in abandoned biomass; Described allotment, refer to according to the component content such as albumen, polysaccharide in different waste biomass materials, by a certain percentage proportioning raw material, and the supplementary of correspondence adds the fertilizer such as nitrogen, phosphorus, potassium, reach be suitable for fermentable growth needed for nutrition and element proportioning; The interpolation of described trace element, refers to the needs according to target chlorella product, and corresponding adds zinc, chromium (Cr ³⁺), the inorganic salt element of the trace such as selenium, iodine.

3. organic chlorella production method of a kind of rich trace elements according to claim 1, it is characterized in that: producing of fertilizer anaerobic fermented liquid, cellulose-decomposing bacterium and proteolysis bacterium bacterial classification will be added in microbiological anaerobic fermention medium, under anaerobic ferment, obtain containing organic fermented liquids such as monose, oligosaccharides, low first organic acid, total free aminoacids, protein peptide, thalline, in fermented liquid, be rich in zinc, chromium (Cr simultaneously ³⁺), the trace element such as selenium, iodine; Described fertilizer anaerobically fermenting, under referring to oxygen free condition, the process of cellulose-decomposing bacterium, amylolysis bacterium, steatolysis bacterium and proteolysis bacterium is cultivated in microbiological anaerobic fermention medium, its principle utilizes cellulose-decomposing bacterium, and in the process of amylolysis bacterium and steatolysis bacterium growth and breeding in the medium, the material such as Mierocrystalline cellulose, starch, fat decomposed in substratum becomes monose, oligosaccharides and the low first organic acid etc. that other mushrooms or chlorella can directly utilize; Utilize proteolysis bacterium, in the process of growth and breeding in the medium, decomposition of protein class material, if the material such as thalline, albumen is the process of the chlorella total free aminoacids that can directly utilize and protein small peptide;

Further, described cellulose-decomposing bacterium, is referred to and can under anaerobic, be produced and metabolism by thalline, Mierocrystalline cellulose in decomposing biomass is the bacterial classification of low first carbohydrate such as monose, oligosaccharides and formic acid, acetic acid, propionic acid, butyric acid, as each kind of strain of Clostridium;

Further, described amylolysis bacterium, is referred to and can under anaerobic, be produced and metabolism by thalline, starch in decomposing biomass is the bacterial classification of low first carbohydrate such as monose, oligosaccharides and formic acid, acetic acid, propionic acid, butyric acid, as some of Bacillus subtilus plant strain;

Further, described steatolysis bacterium, is referred to and can under anaerobic, be produced and metabolism by thalline, fat in decomposing biomass is the bacterial classification of low first carbohydrate such as monose, oligosaccharides and formic acid, acetic acid, propionic acid, butyric acid, as Clostridium and Bacillus subtilus;

Further, described protein decomposing bacteria, refer to and can under anaerobic, be produced and metabolism by thalline, the protein in decomposing biomass is the bacterial classification of total free aminoacids and protein small peptide, and in environment, most anerobe all has protelytic function.

4. organic chlorella production method of a kind of rich trace elements according to claim 1, it is characterized in that: organic chlorella produces, by the fermented liquid sterilizing of the rich trace elements of acquisition, after allotment, be added into chlorella Cultivation container, carry out chlorella cultivation; After chlorella has cultivated, centrifugally to gather, dry, namely obtain the organic chlorella product being enriched all kinds of trace element, can develop for follow-up further processing; Described chlorella cultivation, refer to and utilize the fermented liquid being rich in monose, oligosaccharides, low first organic acid, total free aminoacids, protein small peptide and each trace element obtained as substratum, as the raw material that chlorella is cultivated, in culture vessel, cultivate the process of chlorella; Further, what chlorella culture vessel referred to must be fermentor tank, Closed photobioreactor or open pond, and the series combination of above different vessels; Described centrifugally to gather, refer to the process utilizing the centrifugal chlorella of gathering of disc centrifuge to obtain chlorella underflow; Described drying, refers to and utilizes spray-drying tower dried pellet algae underflow, obtains the process of chlorella dry powder.