Deficiency in the glycerol channel Fps1p confers increased freeze tolerance to yeast cells: application of the fps1? mutant to frozen dough technology

2004 ◽  
Vol 66 (3) ◽  
pp. 303-305 ◽  
Author(s):  
Shingo Izawa ◽  
Kayo Ikeda ◽  
Kazuhiro Maeta ◽  
Yoshiharu Inoue
Author(s):  
Н.Н. АЛЕХИНА ◽  
Е.И. ПОНОМАРЕВА

Для обогащения хлебобулочных изделий используют биоактивированное зерно пшеницы, что обусловливает введение дополнительной стадии технологического процесса – биоактивации. Снизить трудоемкость и сократить продолжительность процесса приготовления зернового хлеба из биоактивированной пшеницы, что особенно важно в условиях предприятий малой мощности, позволяет применение технологии глубокой заморозки теста. Однако продолжительная заморозка снижает активность клеток бродильной микрофлоры теста, поэтому необходимо установить оптимальный срок хранения замороженного тестового полуфабриката. Для исследований зерно пшеницы предварительно очищали от сорной и зерновой примеси, мыли, оставляли для набухания в воде на 22–24 ч, проращивали в течение 11–12 ч и измельчали путем пропускания через матрицу с диаметром отверстий 2 мм. Тестовые полуфабрикаты изготавливали по рецептуре для хлеба «Айсбрэд», затем их подвергали шоковой заморозке при температуре –35°С и хранили при –18°С в течение 7, 14, 21 и 28 сут. Перед использованием тесто размораживали, полуфабрикат выбраживал, из него формовали тестовые заготовки массой 0,07 кг, которые после расстойки выпекали. Контролем служило тесто без заморозки, приготовленное по рецептуре для хлеба «Колосок». Через 20 ч после выпечки оценивали физико-химические и органолептические показатели изделий. Методом микроскопирования выявлено, что с увеличением продолжительности хранения в замороженных тестовых полуфабрикатах повышается содержание мертвых клеток дрожжей. Наименьшее количество их наблюдалось в образце теста без заморозки – 12,2%, что на 29,0% меньше, чем в образце замороженного теста при хранении 28 сут. Установлено, что при хранении замороженного теста в течение 7, 21 и 28 сут содержание восстановленного глутатиона в нем соответственно на 1,6; 7,4 и 7,8% выше, чем в тесте без заморозки. Количество выделенного диоксида углерода через 120 мин брожения в образцах замороженного теста, хранившихся 7, 14, 21 и 28 сут, было меньше в 1,4; 1,5; 2,5 и 2,8 раза соответственно, чем в контрольном образце. По показателям качества изделия, выпеченные из теста после заморозки и без нее, существенно не отличались. На основании результатов исследований рекомендовано хранить замороженный тестовый полуфабрикат для приготовления зернового хлеба при температуре –18°С не более чем 21 сут. For the enrichment of bakery products, bioactivated wheat grain is used, which leads to the introduction of an additional stage of the technological process – bioactivation. The use of deep-freezing technology allows you to reduce the labor intensity and reduce the duration of the process of preparing grain bread from bioactivated wheat, which is especially important in the conditions of small-scale enterprises. However prolonged freezing reduces the activity of the cells of the fermentation microflora of the dough, so it is necessary to establish the optimal shelf life of the frozen dough semi-finished product. For research, wheat grain was previously cleaned from weed and grain impurities, washed, left to swell in water for 22–24 hours, germinated for 11–12 hours and ground by passing through a matrix with a hole diameter of 2 mm. The dough pieces were made according to the recipe for «Aysbred» bread, then they were subjected to shock freezing at a temperature of –35°C and stored at –18°C for 7, 14, 21 and 28 days. Before use, the dough was thawed, the semi-finished product was fermented, and dough pieces weighing 0,07 kg were formed from it, which were baked after proofing. The control was the dough without freezing, prepared according to the recipe for bread «Kolosok». After 20 hours after baking, the physico-chemical and organoleptic parameters of the baked bread were evaluated. Microscopy revealed that with an increase in the shelf life of frozen semi-finished products, the content of dead yeast cells increases. Their lowest number was observed in the sample of the test without freezing – 12,2%, which is by 29,0% less than in the sample of the frozen test, stored for 28 days. It was found that in the frozen dough samples stored for 7, 21 and 28 days, the content of reduced glutathione was by 1,6; 7,4 and 7,8% higher respectively, than in the control sample. After 120 min of fermentation, the amount of carbon dioxide released in the frozen dough samples stored for 7, 14, 21 and 28 days was in 1,4; 1,5; 2,5 and 2,8 times less respectively, than in the test without freezing. In terms of quality, the products baked from the dough after freezing and without it did not differ significantly. Based on the research results, it is recommended to store the frozen test semi-finished product for the preparation of grain bread at a temperature of –18°C for no more than 21 days.


2004 ◽  
Vol 70 (6) ◽  
pp. 3377-3382 ◽  
Author(s):  
An Tanghe ◽  
Patrick Van Dijck ◽  
Didier Colavizza ◽  
Johan M. Thevelein

ABSTRACT Previous observations that aquaporin overexpression increases the freeze tolerance of baker's yeast (Saccharomyces cerevisiae) without negatively affecting the growth or fermentation characteristics held promise for the development of commercial baker's yeast strains used in frozen dough applications. In this study we found that overexpression of the aquaporin-encoding genes AQY1-1 and AQY2-1 improves the freeze tolerance of industrial strain AT25, but only in small doughs under laboratory conditions and not in large doughs under industrial conditions. We found that the difference in the freezing rate is apparently responsible for the difference in the results. We tested six different cooling rates and found that at high cooling rates aquaporin overexpression significantly improved the survival of yeast cells, while at low cooling rates there was no significant effect. Differences in the cultivation conditions and in the thawing rate did not influence the freeze tolerance under the conditions tested. Survival after freezing is determined mainly by two factors, cellular dehydration and intracellular ice crystal formation, which depend in an inverse manner on the cooling velocity. In accordance with this so-called two-factor hypothesis of freezing injury, we suggest that water permeability is limiting, and therefore that aquaporin function is advantageous, only under rapid freezing conditions. If this hypothesis is correct, then aquaporin overexpression is not expected to affect the leavening capacity of yeast cells in large, industrial frozen doughs, which do not freeze rapidly. Our results imply that aquaporin-overexpressing strains have less potential for use in frozen doughs than originally thought.


1996 ◽  
Vol 60 (11) ◽  
pp. 1874-1876 ◽  
Author(s):  
Yoko Murakami ◽  
Kumio Yokoigawa ◽  
Fusako Kawai ◽  
Hiroyasu Kawai

2003 ◽  
Vol 69 (1) ◽  
pp. 212-219 ◽  
Author(s):  
Yuko Morita ◽  
Shigeru Nakamori ◽  
Hiroshi Takagi

ABSTRACT We previously isolated a mutant which showed a high tolerance to freezing that correlated with higher levels of intracellular l-proline derived from l-proline analogue-resistant mutants. The mutation responsible for the analogue resistance and l-proline accumulation was a single nuclear dominant mutation. By introducing the mutant-derived genomic library into a non-l-proline-utilizing strain, the mutant was found to carry an allele of the wild-type PRO1 gene encoding γ-glutamyl kinase, which resulted in a single amino acid replacement; Asp (GAC) at position 154 was replaced by Asn (AAC). Interestingly, the allele of PRO1 was shown to enhance the activities of γ-glutamyl kinase and γ-glutamyl phosphate reductase, both of which catalyze the first two steps of l-proline synthesis from l-glutamate and which together may form a complex in vivo. When cultured in liquid minimal medium, yeast cells expressing the mutated γ-glutamyl kinase were found to accumulate intracellular l-proline and showed a prominent increase in cell viability after freezing at −20°C compared to the viability of cells harboring the wild-type PRO1 gene. These results suggest that the altered γ-glutamyl kinase results in stabilization of the complex or has an indirect effect on γ-glutamyl phosphate reductase activity, which leads to an increase in l-proline production in Saccharomyces cerevisiae. The approach described in this paper could be a practical method for breeding novel freeze-tolerant yeast strains.


2008 ◽  
Vol 74 (18) ◽  
pp. 5845-5849 ◽  
Author(s):  
Tomohiro Kaino ◽  
Tetsuya Tateiwa ◽  
Satomi Mizukami-Murata ◽  
Jun Shima ◽  
Hiroshi Takagi

ABSTRACT We constructed self-cloning diploid baker's yeast strains by disrupting PUT1, encoding proline oxidase, and replacing the wild-type PRO1, encoding γ-glutamyl kinase, with a pro1(D154N) or pro1(I150T) allele. The resultant strains accumulated intracellular proline and retained higher-level fermentation abilities in the frozen doughs than the wild-type strain. These results suggest that proline-accumulating baker's yeast is suitable for frozen-dough baking.


2007 ◽  
pp. 293-301 ◽  
Author(s):  
Dusanka Pejin ◽  
Irena Dosanovic ◽  
Stevan Popov ◽  
Zvonimir Suturovic ◽  
Jovana Rankovic ◽  
...  

The need to freeze dough is increasing in bakery production. Frozen dough can be stored for a long time without quality change. The capacity of bakery production can be increased in this way, and in the same time, the night shifts can be decreased. Yeast cells can be damaged by freezing process resulting in poor technological quality of dough after defrostation (longer fermentation of dough). The influence of frozen storage time of dough on survival percentage of Saccharomyces cerevisiae was investigated. Dough samples were taken after 1, 7, 14 and 28 days of frozen storage at -20?C. After defrosting, at room temperature, samples were taken from the surface and the middle part of dough (under aseptic conditions), and the percentage of living S. cerevisiae cells was determined. During frozen storage of dough, the number of living S. cerevisiae decreased. After 28 days of frozen storage, the percentage of live cells on the surface and inside the dough was 53,1% and 54,95%, respectively. The addition of k-carragenan to dough increased the percentage of living cells in the middle part of dough up to 64,63%. Pure cultures, isolated from survived S. cerevisia cells in frozen dough by agar plates method (Koch's method), were multiplied in optimal liquid medium for yeasts. The content of cytochromes in S. cerevisiae cells was determined by spectrophotometric method. The obtained results showed that the content of cytochromes in survived S. cerevisiae cells was not affected by dough freezing process. Growth rate and fermentative activity (Einchor's method) were determined in multiplied cells.


2002 ◽  
Vol 68 (12) ◽  
pp. 5981-5989 ◽  
Author(s):  
An Tanghe ◽  
Patrick Van Dijck ◽  
Françoise Dumortier ◽  
Aloys Teunissen ◽  
Stefan Hohmann ◽  
...  

ABSTRACT Little information is available about the precise mechanisms and determinants of freeze resistance in baker's yeast, Saccharomyces cerevisiae. Genomewide gene expression analysis and Northern analysis of different freeze-resistant and freeze-sensitive strains have now revealed a correlation between freeze resistance and the aquaporin genes AQY1 and AQY2. Deletion of these genes in a laboratory strain rendered yeast cells more sensitive to freezing, while overexpression of the respective genes, as well as heterologous expression of the human aquaporin gene hAQP1, improved freeze tolerance. These findings support a role for plasma membrane water transport activity in determination of freeze tolerance in yeast. This appears to be the first clear physiological function identified for microbial aquaporins. We suggest that a rapid, osmotically driven efflux of water during the freezing process reduces intracellular ice crystal formation and resulting cell damage. Aquaporin overexpression also improved maintenance of the viability of industrial yeast strains, both in cell suspensions and in small doughs stored frozen or submitted to freeze-thaw cycles. Furthermore, an aquaporin overexpression transformant could be selected based on its improved freeze-thaw resistance without the need for a selectable marker gene. Since aquaporin overexpression does not seem to affect the growth and fermentation characteristics of yeast, these results open new perspectives for the successful development of freeze-resistant baker's yeast strains for use in frozen dough applications.


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