Method for producing cream-aroma selenium-enriched bread from yeast display lipase
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A technology of lipase and rice lipase, which is applied in the field of food processing, can solve the problems of poor stability of lipase, high price, and the successful display of rice lipase by yeast.
Inactive Publication Date: 2014-04-23
CHANGSHA UNIVERSITY OF SCIENCE AND TECHNOLOGY
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[0003] At present, on the one hand, due to poor stability and high price, lipase mainly relies on imports in my country, which limits the large-scale use of lipase; Rice lipase is difficult to separate from rice, and it has not been practically used, and it has been discarded as a harmful substance due to inactivation of the enzyme.
[0004] In recent years, the rapid development of modern biotechnology has brought the development and utilization of lipase to a new level, such as the doctoral thesis (Liu Wenshan. Research on the surface display of lipase in Saccharomyces cerevisiae [D] Wuhan, Huazhong University of Science and Technology, 2010 ), the surface display of lipase on yeast has the advantages of increasing the yield of lipase and not requiring separation and purification during use. Report on Rice Lipase and Its Application
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Embodiment 1
[0013] 1. Pichia pastoris displays rice lipase
[0014] 1. The Rlipase gene synthesis was entrusted to a biological company (in this case, Nanjing GenScript Biotechnology Co., Ltd.), and the gene was cloned on the pUC57 vector (pUC57-Rlipase). The DNA sequence is shown in SEQ1.
[0015] 2. Rlipase was subcloned into pPIC9K vector
[0016] The pUC57-Rlipase and pPIC9K vectors were digested with Eco RI and Not I respectively, and the enzyme digestion reaction systems were as follows:
[0017] pUC57-Rlipase plasmid 34 ml
[0018] 10* buffer 4ml
[0019] Eco RI 1ml
[0020] Not I 1ml
[0021] A total of 40ml system,
[0022] pPIC9K plasmid 6 ml
[0023] 10* buffer 2ml
[0024] Eco RI 1ml
[0025] Not I 1ml
[0026] wxya 2 O 10ml
[0027] 20 ml system in total,
[0028] Mix them separately and place them in a water bath at 37°C for 4 h, then identify them by 1.2% agarose gel electrophoresis, the results are as follows: figure 1 shown.
[0029] 3. Recover the target ge...
Embodiment 2
[0073] 1. Pichia pastoris displays rice lipase: the same as in Example 1.
[0074] Two, selenium-enriched culture of baker's yeast: same as embodiment 1.
[0075] 3. Production of frankincense selenium-enriched bread:
[0076] as attached image 3 As shown, add 3 parts (wet weight) of the yeast displaying rice lipase expanded and cultured in the above 1 and 4) to 400 parts of milk fat (30% fat (w / w)) at 50 ° C After 4 hours of fat esterification reaction, add 1500 parts of flour, then add 3 parts of selenium-enriched yeast (selenium content 1582.31 μg / g) and 4 parts of salt, 40 parts of white sugar, appropriate amount Water and dough for 20 minutes, proofed for 40 minutes, shaped and baked to obtain delicious health-care bread rich in frankincense and rich in organic selenium.
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Abstract
The invention discloses a method for producing cream-aroma selenium-enriched bread from yeast display lipase. The method includes displaying rice lipase on the surface of pichia pastoris, performing multiplication culture for 48-96 hours, enabling yield of the rice lipase to sharply increase with yield of yeast, adding 1-4 parts of saccharomycetes displayed with the rice lipase into 200-300 parts of cream liquid for esterlysis reaction at the temperature of 35-55 DEG C for 2-6 hours, subjecting the mixture to filtering separation, washing the yeast to allow the same to be reusable, adding filtrate into 1000 parts, by weight, of flour, then adding 1-4 parts of selenium-enriched yeast and other raw materials, and performing dough kneading, fermentation, forming and baking to obtain the delicious healthcare bread with rich cream aroma and rich organic selenium.
Description
technical field [0001] The invention relates to the technical field of food processing, in particular to a method for producing frankincense and selenium-enriched bread by using lipase displayed on the surface of yeast to esterify fat to enhance flavor and enriching yeast with selenium. Background technique [0002] Selenium is an essential trace element for the human body. 72% of my country's areas are selenium-deficient or low-selenium areas, which seriously threaten people's health. The development of bread rich in frankincense and safe supplementation of organic selenium can effectively alleviate this phenomenon, and lipase Fat esterification and yeast selenium enrichment are effective means of enriching selenium for flavor enhancement. [0003] At present, on the one hand, due to poor stability and high price, lipase mainly relies on imports in my country, which limits the large-scale use of lipase; Rice lipase is difficult to separate from rice, and it has not been prac...
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