CN101870960A - Lactobacillus acidophilus composite antifreeze factor and freeze-drying method - Google Patents
Lactobacillus acidophilus composite antifreeze factor and freeze-drying method Download PDFInfo
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- CN101870960A CN101870960A CN 201010207358 CN201010207358A CN101870960A CN 101870960 A CN101870960 A CN 101870960A CN 201010207358 CN201010207358 CN 201010207358 CN 201010207358 A CN201010207358 A CN 201010207358A CN 101870960 A CN101870960 A CN 101870960A
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Abstract
The invention provides a lactobacillus acidophilus composite antifreeze factor and a freeze-drying method. According to a formula, 1 liter of the composite antifreeze factor comprises 2 to 6mL of glycerine, 2 to 8g of sodium chloride, 0 to 4g of trehalose and 0 to 0.8g of alanine. The method comprises the following steps of: adding the composite antifreeze factor into a culture medium for culturing the lactobacillus acidophilus; centrifuging and then collecting the lactobacillus acidophilus, cleaning the lactobacillus acidophilus by using normal saline, centrifuging and then removing clear liquid; and preparing the lactobacillus acidophilus into lactobacillus acidophilus suspension by using the normal saline, then uniformly mixing the lactobacillus acidophilus suspension with a protective agent and finally performing freeze drying. The freeze-drying antifreeze factor of the invention is directly added into the culture medium to prepare lactobacillus acidophilus powder; after freeze drying, the maximum survival rate of the lactobacillus acidophilus reaches 82.75 percent and a powder activity reaches over 1*10<11>cfu/g; and the survival rate of the lactobacillus acidophilus is over three times the survival rate (40.67 percent) of the lactobacillus acidophilus powder prepared without the antifreeze factor for the freeze drying.
Description
Technical field
The present invention relates to a kind of lactobacillus acidophilus composite antifreeze factor and freeze-drying method.
Background technology
Lactobacterium acidophilum is very to pay attention to one of probiotic bacterium of research and development in the present milk-acid bacteria family, also is one of the main bacteria seed of the probiotics of domestic and international application.It extensively is distributed in the animals and plants goods of digestive tube, vagina and the milk-product of humans and animals body and fermentation, is one of important microbe in the human intestinal.Lactobacterium acidophilum has microbial ecological balance, promoting digestion, cancer-resisting, strengthening immunity, the anti-ageing function of waiting for a long time in the human body of keeping, and its effect in multiple disease treatment is remarkable.Lactobacillus acidophilus high activity bacterium powder has been widely used in protective foods, and in the industries such as medicine and food, therefore, preparation high reactivity bacterium powder has wide commercial promise.
Vacuum Freezing ﹠ Drying Technology is that modern industry prepares high reactivity bacterium powder method commonly used.Yet, can damage microorganism cells in the vacuum lyophilization process, for example, the eutectic damage that the inside and outside solution of cell injury that ice crystal causes and cell causes.Eutectic is meant that the part (water and solute) do not freezed in the solution is simultaneously freezing and is the phenomenon of solid (hydrate).Kristiansen discovers when the liposome in the salts solution freezes that the appearance of eutectic obviously increases the liposome degree of injury.Eutectic phenomena not only cell membrane produces physical abuse, simultaneously also can the interior active substance of damaging cells.
Studies show that: in the lyophilize freeze-drying process, add adequate protective agent and can effectively protect thalline, improve the thalline survival rate.But the increase of protective material consumption must reduce the activity of bacterium powder.Therefore, improve the anti-refrigerating capacity of thalline itself and be only the key that addresses this problem.Add the composition (small-molecule substance enters in the enchylema, and macromolecular substance sticks on the cytolemma) that some materials can change enchylema and cytolemma in fermented liquid, the generation of eutectic phenomena in the freezing-inhibiting drying process effectively improves thalline hole refrigerating capacity.
Summary of the invention
Technical problem to be solved by this invention provides a kind of lactobacillus acidophilus composite antifreeze factor and freeze-drying method, adds this compound anti-freezing factor and cultivate the Lactobacterium acidophilum that obtains in substratum, and its survival rate can improve more than the twice.
For achieving the above object, the invention provides a kind of lactobacillus acidophilus composite antifreeze factor, its prescription consists of: glycerine 2-6mL/L, sodium-chlor 2-8g/L, trehalose 0-4g/L, L-Ala 0-0.8g/L.
For achieving the above object, the present invention also provides a kind of Lactobacterium acidophilum freeze-drying method: the compound anti-freezing factor is provided, consists of: glycerine 2-6mL/L, sodium-chlor 2-8g/L, trehalose 0-4g/L, L-Ala 0-0.8g/L; This composite frozen factor is added in the aseptic MRS substratum cultivates Lactobacterium acidophilum; Behind the centrifugal collection thalline, clean thalline with physiological saline; After with physiological saline thalline being mixed with bacteria suspension, be to mix at 1: 3 according to volume ratio with bacteria suspension and protective material; Carry out lyophilize at last.
The present invention compared with prior art, at least have the following advantages: after in substratum, adding the compound anti-freezing factor of the present invention, can make the survival rate of Lactobacterium acidophilum in freezing dry process up to 82.75%, if should compoundly not resist freezing factor and add, then the survival rate of thalline in freezing dry process only 40.67%, therefore, in substratum, add the anti-freezing factor of the present invention, the survival rate of thalline in freezing dry process can be improved more than the twice.
Embodiment
The compound anti-freezing factor of millipore filtration degerming being added in the aseptic MRS substratum, cultivate Lactobacterium acidophilum 24h, is that centrifugal 10min collects thalline under the condition of 6000r/min at rotating speed, subsequently, cleans thalline 2 times with physiological saline, centrifugal abandoning supernatant; With physiological saline thalline is mixed with 8 ± 0.5 * 10
10The bacteria suspension of cfu/mL, according to volume ratio be 1: 3 with bacteria suspension and protective material solution (skim-milk 100g/L, sucrose 5g/L, xitix 0.1g/L, the aqueous solution of sodium-chlor 8.5g/L) mix, be loaded in the freeze-drying pipe, dress sample height is 0.3-0.5cm, under the room temperature behind the static 30min, after putting into-35 ℃ of refrigerator pre-freeze 24h, putting into vacuum freeze drier, is-56 ℃ in temperature, and vacuum tightness is freeze-drying 24h under the condition of 4-6Pa; Redissolve with physiological saline, MRS solid medium counting is measured thalline survival rate and bacterium powder activity.
The anti-freezing factor of adding different proportionings is cultivated checking result such as the table 1 that the thalline that obtains carries out freeze dried survival rate in fermented liquid:
The thalline that the compound anti-freezing factor fermentation of the different proportionings of table 1 interpolation obtains carries out the checking result of freeze dried survival rate
| The compound anti-freezing factor | Survival rate |
| Glycerine 4mL/L+ sodium-chlor 4g/L+ trehalose 4g/L+Ala 0.4g/L | ??82.75％ |
| Glycerine 6mL/L+ sodium-chlor 8g/L+ trehalose 4g/L+Ala 0.6g/L | ??75.32％ |
| Glycerine 6mL/L+ sodium-chlor 6g/L+ trehalose 2g/L+Ala 0.3g/L | ??73.62％ |
| Glycerine 2mL/L+ sodium-chlor 4g/L+ trehalose 4g/L+Ala 0.8g/L | ??67.25％ |
| Glycerine 3mL/L+ sodium-chlor 4g/L+ trehalose 2g/L+Ala 0.3g/L | ??64.00％ |
| Glycerine 2mL/L+ sodium-chlor 2g/L+ trehalose 2g/L+Ala 0.4g/L | ??62.02％ |
| Glycerine 4mL/L+ sodium-chlor 4.5g/L | ??60.73％ |
| Do not add the compound anti-freezing factor | ??40.67％ |
By above table as can be known, when not adding the compound anti-freezing factor in the fermented liquid, freeze-drying prepares the bacterium powder, its thalline survival rate only 40.67%; And the compound anti-freezing factor of interpolation the present invention in the fermented liquid, freeze-drying prepares the bacterium powder, the survival rate of its thalline is at least more than 60.73%, wherein, when it consists of glycerine: 4mL/L, sodium-chlor: 4g/L, trehalose: 4g/L, L-Ala: during 0.4g/L, its thalline survival rate is the highest, reach 82.75%, compare with not adding the compound anti-freezing factor, it is many to have improved twice, cuts bacterium powder activity and can reach 1 * 10
11More than the cfu/g.
The above only is one embodiment of the present invention, it or not whole or unique embodiment, the conversion of any equivalence that those of ordinary skills take technical solution of the present invention by reading specification sheets of the present invention is claim of the present invention and contains.
Claims (10)
1. lactobacillus acidophilus composite antifreeze factor, it is characterized in that: its prescription consists of: glycerine 2-6mL/L, sodium-chlor 2-8g/L, trehalose 0-4g/L, L-Ala 0-0.8g/L.
2. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 4mL/L, sodium-chlor 4g/L, trehalose 4g/L, L-Ala 0.4g/L.
3. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 6mL/L, sodium-chlor 8g/L, trehalose 4g/L, L-Ala 0.6g/L.
4. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 6mL/L, sodium-chlor 6g/L, trehalose 2g/L, L-Ala 0.3g/L.
5. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 2mL/L, sodium-chlor 4g/L, trehalose 4g/L, L-Ala 0.8g/L.
6. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 3mL/L, sodium-chlor 4g/L, trehalose 2g/L, L-Ala 0.3g/L.
7. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 2mL/L, sodium-chlor 2g/L, trehalose 2g/L, L-Ala 0.4g/L.
8. lactobacillus acidophilus composite antifreeze factor as claimed in claim 1 is characterized in that: its prescription consists of: glycerine 4mL/L, sodium-chlor 4.5g/L.
9. a Lactobacterium acidophilum freeze-drying method is characterized in that: the compound anti-freezing factor is provided, consists of: glycerine 2-6mL/L, sodium-chlor 2-8g/L, trehalose 0-4g/L, L-Ala 0-0.8g/L; This composite frozen factor is added in the aseptic MRS substratum cultivates Lactobacterium acidophilum; Behind the centrifugal collection thalline, clean thalline with physiological saline; After with physiological saline thalline being mixed with bacteria suspension, be to mix at 1: 3 according to volume ratio with bacteria suspension and protective material; Carry out lyophilize at last.
10. Lactobacterium acidophilum freeze-drying method as claimed in claim 9 is characterized in that: described protectant component is: skim-milk 100g/L, sucrose 5g/L, xitix 0.1g/L, sodium-chlor 8.5g/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102408992A (en) * | 2011-11-23 | 2012-04-11 | 陕西农产品加工技术研究院 | Lactobacillus acidophilus anti-freeze culture medium and application method thereof |
| CN102978150A (en) * | 2012-11-19 | 2013-03-20 | 陕西科技大学 | Streptococcus thermophilus composite antifreezing factor |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101486986A (en) * | 2009-02-12 | 2009-07-22 | 上海谱莱生物技术有限公司 | Preparation of freeze-dried Lactobacillus acidophilus powder |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101486986A (en) * | 2009-02-12 | 2009-07-22 | 上海谱莱生物技术有限公司 | Preparation of freeze-dried Lactobacillus acidophilus powder |
Non-Patent Citations (2)
| Title |
|---|
| 《现代食品科技》 20050301 蒲丽丽等 乳酸菌冻干保护剂及保护机理的研究进展 第148页 1－10 第21卷, 第1期 2 * |
| 《食品工业科技》 20070525 徐丽萍 嗜酸乳杆菌冻干菌粉保护剂选择的研究 第119－121页 1－10 第28卷, 第5期 2 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102408992A (en) * | 2011-11-23 | 2012-04-11 | 陕西农产品加工技术研究院 | Lactobacillus acidophilus anti-freeze culture medium and application method thereof |
| CN102978150A (en) * | 2012-11-19 | 2013-03-20 | 陕西科技大学 | Streptococcus thermophilus composite antifreezing factor |
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