CN110623206A - Preparation method of lovastatin-rich functional red yeast rice - Google Patents
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Abstract
The invention discloses a preparation method of lovastatin-rich functional red yeast rice, which comprises the steps of rice soaking, high-temperature sterilization, seed liquid preparation, inoculation, fermentation and drying, and is characterized in that: in the fermentation step, the fermentation culture time is controlled to be 18-24 days, cooling fermentation is adopted, the culture temperature is controlled to be 30-35 ℃ at 0-5 days during the fermentation culture period, and the culture temperature is controlled to be 22-27 ℃ from 6d to the end of fermentation during the fermentation culture period; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with water and controlling humidity by using fermentation supernatant, and the rest time is supplemented with sterile water and controlling humidity; turning over the yeast every two days after the 16 th day of the fermentation culture until the fermentation is finished. The preparation method can improve the lovastatin content in the functional red yeast rice and improve the fermentation effect and the product quality of the functional red yeast rice.
Description
Technical Field
The invention belongs to the technical field of microbial solid state fermentation, and particularly relates to a preparation method of lovastatin-rich functional red yeast rice.
Background
Red yeast rice/powder is a medicinal and edible product prepared by fermenting red yeast rice with traditional large grains as raw materials, is a traditional color symbol in China, and has a history of thousands of years. The production of red yeast rice is mainly concentrated in south-east Asia countries, has the functions of regulating blood fat and blood pressure, enhancing immunity, increasing antibodies and the like, and is quite common in the field of food industry. The Chinese red yeast rice is mainly eaten in provinces and cities of Jiangsu, Shanghai, Fujian, Jiangxi, Zhejiang, Shandong and the like, and the red yeast rice is eaten by people at a rate of 14-80 g/d. Red yeast products on the market are mainly represented in pigment red yeast (containing pigment for coloring) and functional red yeast (containing monacolin K, also called lovastatin for health care and medicine).
The chemical structure of the open-ring hydroxy acid part in the chemical structure of the lovastatin in the functional red yeast rice is very similar to that of 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA), so that the lovastatin open-ring hydroxy acid part can competitively inhibit a rate-limiting enzyme HMG-CoA reductase in the cholesterol synthesis process, and the functional red yeast rice becomes a pure natural product with the functions of reducing blood fat and cholesterol. Secondly, the functional red yeast rice contains ergosterol, gamma-aminobutyric acid, monascus pigment, coenzyme Q10, rich enzymes and other bioactive substances, and is a natural, nutritional, health-care, medicinal and safe health-care food and medicinal raw material.
At present, the lipid-lowering medicines in the market mainly utilize red yeast rice as raw material components, such as 'Zhidakang' of Beidaweixin and 'Zhibituo' of Chengdouao, and occupy leading positions in domestic lipid-regulating medicines. With the upgrading of consumption, the diversification of products and the like rise, and a new product of the baby haha red yeast rice quinoa meal biscuit is published by a new product issuing society (2019) in Hangzhou; the Hubei algae Shanghai Biotechnology Limited announces (2019) that the first red rice liquid fermented drink-MONABABABY red rice baby is on the market, develops red rice red energy drinks, red rice ferment drinks, red rice composite drinks and other series drinks, and enters Guangdong area. The data shows that the demand of the Chinese functional red yeast rice is 439 tons in 2016, and the year is increased by 9.3 percent.
The production of functional red yeast rice still mainly uses the solid state fermentation technology of rice as a substrate, and is divided into bottle type fermentation, tray fermentation, fermentation in a yeast pool and solid state fermentation tank fermentation according to the type of a container. However, the production technology of a large-scale fermentation tank is not realized at an industrial level because of the specific growth metabolism and solid state fermentation technology of monascus, functional monascus is mainly traditional bottle fermentation, but the lovastatin content level is in the stage of 4 mg/g-30 mg/g, the price of the functional monascus is greatly different along with the lovastatin content, and it is necessary to develop a novel solid state fermentation technology or breed monascus with stable and high lovastatin yield to improve the lovastatin content in the functional monascus.
Disclosure of Invention
In order to solve the problems that the lovastatin content is generally low and the like in the functional red yeast rice produced by the traditional solid state fermentation technology taking bulk grain rice as a substrate, the invention provides a preparation technology for preparing the functional red yeast rice rich in lovastatin.
The invention is realized by the following technical scheme.
A preparation method of lovastatin-rich functional red yeast rice comprises the steps of rice soaking, high-temperature sterilization, seed liquid preparation, inoculation, fermentation and drying, and is characterized in that:
in the fermentation step, the fermentation culture time is controlled to be 18-24 days, cooling fermentation is adopted, the culture temperature is controlled to be 30-35 ℃ at 0-5 days during the fermentation culture period, and the culture temperature is controlled to be 22-27 ℃ from 6d to the end of fermentation during the fermentation culture period; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with water and controlling humidity by using fermentation supernatant, and the rest time is supplemented with sterile water and controlling humidity; turning over the yeast every two days after the 16 th day of the fermentation culture until the fermentation is finished.
As a specific technical scheme, the preparation method of the lovastatin-rich functional red yeast rice comprises the following steps:
1) soaking rice: soaking rice in glacial acetic acid aqueous solution, then fishing out, draining, and collecting for later use;
2) high-temperature sterilization: filling the rice processed in the step (1) into a triangular flask, supplementing water, and performing high-temperature sterilization to prepare a solid fermentation culture medium for later use;
3) preparing a seed solution: activating an monascus test tube, transferring the monascus test tube to a PDA (personal digital Assistant) plate culture medium, adding sterile water, scraping hyphae, and preparing a monascus suspension;
4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2);
5) fermentation: the time of fermentation culture is controlled to be 18-24 d, the culture temperature is controlled to be 30-35 ℃ from 0 th to 5 th d during the fermentation culture period, and the culture temperature is controlled to be 22-27 ℃ from the 6 th d to the end of the fermentation; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with water and controlling humidity by using fermentation supernatant, and the rest time is supplemented with sterile water and controlling humidity; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
(6) and (3) drying: and (5) drying the culture obtained after the culture in the step (5) is finished to obtain the product.
As a specific technical scheme, the rice is indica rice.
As a specific technical scheme, the mass concentration of the glacial acetic acid aqueous solution in the step (1) is 0.1-0.4%.
According to the specific technical scheme, in the step (2), the mass ratio of the supplemented water to the rice is 0.3-0.5: 1.
According to the specific technical scheme, in the step (3), the culture time after the PDA plate culture medium is transferred is 3-5 days, the culture temperature is 30-35 ℃, 8-15 mL of sterile water is added after the monascus purpureus grows in a culture dish with the diameter of 90 mm.
As a specific technical scheme, in the step (4), the inoculation amount of the monascus purpureus strain is 8-12% v/wt of the rice mass.
In the step (6), low-temperature drying is adopted, the drying temperature is controlled to be 40-50 ℃, and the final moisture content of the product is less than 10%.
As a specific technical scheme, the fermentation supernatant is acetic acid fermentation supernatant or lactic acid fermentation supernatant.
The invention has the beneficial effects that:
the preparation method has the advantages of single component of the fermentation medium, simple process operation and improvement on the conventional processes of moistening, moisturizing, turning over and controlling temperature. Specifically, the invention adopts the fermentation supernatant and pure water to moisten water intermittently, on one hand, the humidity control function is achieved; on the other hand, the pH value of the culture environment of the monascus strain is adjusted based on weak acidity of the fermentation liquor, so that the growth and metabolism are facilitated; trace bioactive substances (such as amino acid, organic acid and the like) and metal ions and the like in the fermentation supernatant can regulate the synthetic metabolic pathway of lovastatin; therefore, the content of lovastatin in the functional red yeast rice is improved, the fermentation effect and the product quality of the functional red yeast rice are improved, and the product meets the quality requirement of QB/T2487 plus 2007 (currently effective) in the light industry Standard of the people's republic of China.
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FIG. 1 is a process flow chart of the preparation method of lovastatin-rich functional red yeast rice of the present invention.
Detailed Description
The present invention will be further described with reference to the following detailed description, which should be construed as illustrative only, and not limiting the scope of the invention, which is to be given the full breadth of the appended claims, and all changes that can be made by those skilled in the art and which are, therefore, intended to be embraced therein.
Example 1
A preparation method of lovastatin-rich functional red yeast rice comprises the following steps:
1) soaking rice: the method comprises the following steps of (1) taking indica rice as a raw material, soaking the indica rice in glacial acetic acid water solution with the mass concentration of 0.2% for 4 hours, then fishing out, draining, and collecting for later use;
2) high-temperature sterilization: filling 30g of the indica rice processed in the step (1) into a 250mL triangular flask, then supplementing water, wherein the weight ratio of the supplemented water to the rice is 0.4:1kg/kg, then sterilizing at the high temperature of 121 ℃ for 20min, and then standing and cooling to room temperature to prepare a solid fermentation culture medium for later use;
3) preparing a seed solution: activating an monascus test tube, transferring the activated monascus test tube to a 90mm PDA (personal digital Assistant) plate culture medium for culture, wherein the culture time is 4d, the culture temperature is 32 ℃, adding 10mL of sterile water after the monascus grows over a culture dish, and scraping hyphae to obtain a monascus suspension;
4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2), wherein the inoculation amount is 8% v/wt of the mass of the rice, and then performing fermentation culture;
5) fermentation: controlling the fermentation culture time to be 18 days, and adopting cooling fermentation, wherein the culture temperature is controlled to be 35 ℃ from 0 th to 5 th days during the fermentation culture period, and the culture temperature is controlled to be 27 ℃ from 6 th to 18 th days during the fermentation culture period; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using lactic acid fermentation supernatant; performing secondary yeast turning in the 5 th fermentation culture period, and supplementing water and controlling humidity by using lactic acid fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with lactic acid fermentation supernatant fluid and are controlled humidity, and sterile water is used for supplementing water and controlling humidity in the rest time; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
6) and (3) drying: drying the culture obtained after the culture in the step (5) at a low temperature of 45 ℃ to ensure that the moisture content of the final product is less than 10 percent, thus obtaining the functional red yeast rice; grinding and crushing the functional red yeast rice, and then carrying out quality detection, wherein the content of lovastatin is 37.8 mg/g.
Example 2
A preparation method of lovastatin-rich functional red yeast rice comprises the following steps:
1) soaking rice: the method comprises the following steps of (1) taking indica rice as a raw material, soaking the indica rice in glacial acetic acid water solution with the mass concentration of 0.2% for 4 hours, then fishing out, draining, and collecting for later use;
2) high-temperature sterilization: filling 30g of the indica rice processed in the step (1) into a 250mL triangular flask, then supplementing water, wherein the weight ratio of the supplemented water to the rice is 0.4:1kg/kg, then sterilizing at the high temperature of 121 ℃ for 20min, and then standing and cooling to room temperature to prepare a solid fermentation culture medium for later use;
3) preparing a seed solution: activating an monascus test tube, transferring the activated monascus test tube to a 90mm PDA (personal digital Assistant) plate culture medium for culture, wherein the culture time is 4d, the culture temperature is 32 ℃, adding 10mL of sterile water after the monascus grows over a culture dish, and scraping hyphae to obtain a monascus suspension;
4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2), wherein the inoculation amount is 8% v/wt of the mass of the rice, and then performing fermentation culture;
5) fermentation: controlling the fermentation culture time to be 18 days, and adopting cooling fermentation, wherein the culture temperature is controlled to be 35 ℃ from 0 th to 5 th days during the fermentation culture period, and the culture temperature is controlled to be 27 ℃ from 6 th to 18 th days during the fermentation culture period; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using acetic acid fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using acetic acid fermentation supernatant; turning over the yeast once every 5-16 days of fermentation culture, and supplementing water and controlling humidity, wherein the 12 th and 16 th days are supplemented with acetic acid fermentation supernatant fluid and are controlled humidity, and the rest time is supplemented with sterile water and is controlled humidity; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
6) and (3) drying: drying the culture obtained after the culture in the step (5) at a low temperature of 45 ℃ to ensure that the moisture content of the final product is less than 10 percent, thus obtaining the functional red yeast rice; grinding and crushing the functional red yeast rice, and performing quality detection, wherein the content of lovastatin is 26.5 mg/g.
Example 3
Detection of lovastatin in functional red yeast rice: the Lovastatin standard is Lovastatin (L0790000, sigma), and the content of Lovastatin in the invention is detected according to the content determination method of monacolin K (Monacolin K) in functional red yeast rice in the attachment A of QB/T2487 (national light industry standards for people's republic of China).
Example 4
Preparation of acetic acid fermentation supernatant: acetobacter pasteurianus is subjected to liquid state fermentation (culture medium components are 20g/L of glucose, 10g/L of yeast extract, 0.5g/L of potassium dihydrogen phosphate, 0.5g/L of magnesium sulfate heptahydrate and 20min of sterilization at 121 ℃), the culture temperature is 30 ℃, 150r/min of shake culture is carried out for 3 days, fermentation liquor is centrifuged at 8000r/min at 4 ℃, and supernatant is taken and stored at 4 ℃ for standby.
Example 5
Preparation of lactic acid fermentation supernatant: fermenting the lactobacillus fermentum in liquid state (culture medium components: 40g/L glucose, 5g/L yeast extract, 5g/L peptone, 0.3g/L potassium dihydrogen phosphate, 0.5g/L magnesium sulfate heptahydrate, 0.4g/L zinc sulfate heptahydrate, 40ml/L corn steep liquor, sterilizing at 121 ℃ for 20min), carrying out shake culture at 37 ℃ and 150r/min for 3 days, centrifuging fermentation liquor at 8000r/min at 4 ℃, taking supernatant, and storing at 4 ℃ for later use.
Example 6
Compared with the conventional process, the preparation method of the lovastatin-rich functional red yeast rice
Firstly, preparing lovastatin-rich functional red yeast rice by adopting a conventional process of constant-temperature fermentation and water replenishing and humidity control of a lactic acid aqueous solution, and specifically comprising the following steps of:
1) soaking rice: soaking rice in glacial acetic acid aqueous solution, then fishing out, draining, and collecting for later use;
2) high-temperature sterilization: filling the rice processed in the step (1) into a triangular flask, supplementing water, and performing high-temperature sterilization to prepare a solid fermentation culture medium for later use;
3) preparing a seed solution: activating an monascus test tube, transferring the monascus test tube to a PDA (personal digital Assistant) plate culture medium, adding sterile water, scraping hyphae, and preparing a monascus suspension;
4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2);
5) fermentation: the time of fermentation culture is controlled to be 24d, and the culture temperature is 32 ℃; during the fermentation culture, performing first yeast turning at the 3 rd stage of the fermentation culture, and supplementing water and controlling humidity by using 2% (wt/wt) lactic acid aqueous solution; performing secondary turning over on the fermentation culture at the 5 th day, and supplementing water and controlling humidity by using 2% (wt/wt) lactic acid aqueous solution; turning over the yeast once every 5-16 days after fermentation culture, wherein 2% (wt/wt) lactic acid water solution is used for supplementing water and controlling humidity for 12d and 16d, and sterile water is used for supplementing water and controlling humidity for the rest of time; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
6) and (3) drying: drying the culture obtained after the culture in the step (5) is finished to obtain the product of the invention; grinding and crushing the functional red yeast rice, and then carrying out quality detection, wherein the content of lovastatin is 17.3 mg/g.
Secondly, preparing the lovastatin-rich functional red yeast rice by adopting a conventional process of constant-temperature fermentation and water replenishing and humidity control of an acetic acid aqueous solution, which comprises the following steps:
1) soaking rice: soaking rice in glacial acetic acid aqueous solution, then fishing out, draining, and collecting for later use;
2) high-temperature sterilization: filling the rice processed in the step (1) into a triangular flask, supplementing water, and performing high-temperature sterilization to prepare a solid fermentation culture medium for later use;
3) preparing a seed solution: activating an monascus test tube, transferring the monascus test tube to a PDA (personal digital Assistant) plate culture medium, adding sterile water, scraping hyphae, and preparing a monascus suspension;
4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2);
5) fermentation: the time of fermentation culture is controlled to be 24d, and the culture temperature is 32 ℃; during the fermentation culture, performing first turning over at the 3 rd stage of the fermentation culture, and supplementing water and controlling humidity by using 0.2% (wt/wt) glacial acetic acid aqueous solution; performing secondary turning over at 5d of fermentation culture, and supplementing water and controlling humidity by using 0.2% (wt/wt) glacial acetic acid aqueous solution; turning over the yeast once every 5-16 days after fermentation culture, wherein the 12 th day and the 16 th day are supplemented with 0.2% (wt/wt) glacial acetic acid water solution for moisture control, and the rest time is supplemented with sterile water for moisture control; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
6) and (3) drying: drying the culture obtained after the culture in the step (5) is finished to obtain the product of the invention; grinding and crushing the functional red yeast rice, and then carrying out quality detection, wherein the content of lovastatin is 10.8 mg/g.
Thirdly, comparing the content of the functional red yeast rice lovastatin prepared by the conventional process and the method of the invention
TABLE 1 lovastatin content of functional Red Yeast Rice prepared under different Processes
As can be seen from Table 1, the content of lovastatin in the functional red yeast rice prepared by the method is 26.3-37.8 mg/g, which is far higher than that of lovastatin in the functional red yeast rice prepared by the conventional process by 10.8mg/g and 17.3 mg/g.
Claims (9)
1. A preparation method of lovastatin-rich functional red yeast rice comprises the steps of rice soaking, high-temperature sterilization, seed liquid preparation, inoculation, fermentation and drying, and is characterized in that:
in the fermentation step, the fermentation culture time is controlled to be 18-24 days, cooling fermentation is adopted, the culture temperature is controlled to be 30-35 ℃ at 0-5 days during the fermentation culture period, and the culture temperature is controlled to be 22-27 ℃ from 6d to the end of fermentation during the fermentation culture period; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with water and controlling humidity by using fermentation supernatant, and the rest time is supplemented with sterile water and controlling humidity; turning over the yeast every two days after the 16 th day of the fermentation culture until the fermentation is finished.
2. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 1, characterized by comprising the steps of:
(1) soaking rice: soaking rice in glacial acetic acid aqueous solution, then fishing out, draining, and collecting for later use;
(2) high-temperature sterilization: filling the rice processed in the step (1) into a triangular flask, supplementing water, and performing high-temperature sterilization to prepare a solid fermentation culture medium for later use;
(3) preparing a seed solution: activating an monascus test tube, transferring the monascus test tube to a PDA (personal digital Assistant) plate culture medium, adding sterile water, scraping hyphae, and preparing a monascus suspension;
(4) inoculation: inoculating the bacterial suspension in the step (3) into the solid fermentation culture medium in the step (2);
(5) fermentation: the time of fermentation culture is controlled to be 18-24 d, the culture temperature is controlled to be 30-35 ℃ from 0 th to 5 th d during the fermentation culture period, and the culture temperature is controlled to be 22-27 ℃ from the 6 th d to the end of the fermentation; during the fermentation culture period, performing primary yeast turning in the 3 rd fermentation culture period, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast for the second time in the 5 th step of fermentation culture, and supplementing water and controlling humidity by using fermentation supernatant; turning over the yeast once every 5-16 d of fermentation culture, and supplementing water and controlling humidity, wherein 12d and 16d are supplemented with water and controlling humidity by using fermentation supernatant, and the rest time is supplemented with sterile water and controlling humidity; turning over the yeast every two days after the 16 th day of fermentation culture until the fermentation is finished;
(6) and (3) drying: and (5) drying the culture obtained after the culture in the step (5) is finished to obtain the product.
3. The method of claim 2, wherein said rice is indica rice.
4. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 2, wherein the mass concentration of the glacial acetic acid aqueous solution in the step (1) is 0.1-0.4%.
5. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 2, wherein in the step (2), the supplemented water content and the rice are in a mass ratio of 0.3-0.5: 1 kg/kg.
6. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 2, wherein in the step (3), the culture time after the transfer of the PDA plate culture medium is 3-5 days, the culture temperature is 30-35 ℃, and 8-15 mL of sterile water is added after the monascus overgrowth culture dish with the diameter of 90 mm.
7. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 2, wherein in the step (4), the inoculation amount of the monascus purpureus strain is 8% -12% v/wt of the rice mass.
8. The method for preparing the lovastatin-rich functional red yeast rice as claimed in claim 2, wherein in the step (6), low-temperature drying is adopted, the drying temperature is controlled to be 40-50 ℃, and the final moisture content of the product is less than 10%.
9. The method for preparing lovastatin-rich functional red yeast rice as claimed in any one of claims 1 to 8, wherein the fermentation supernatant is an acetic acid fermentation supernatant or a lactic acid fermentation supernatant.
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CN111328933A (en) * | 2020-03-06 | 2020-06-26 | 山东泰山生力源集团股份有限公司 | Method for improving yolk color and egg quality by using red yeast rice |
CN111407857A (en) * | 2020-03-25 | 2020-07-14 | 杭州天曲生物技术有限公司 | Method for extracting water-soluble red yeast rice |
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CN112779301A (en) * | 2021-03-25 | 2021-05-11 | 安诺康(福建)生物科技有限公司 | Fermentation method for high-yield lovastatin of red yeast rice |
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