JPS6322179A - Production of survivable and acid-resistant dried bifidus cell - Google Patents
Production of survivable and acid-resistant dried bifidus cellInfo
- Publication number
- JPS6322179A JPS6322179A JP61164522A JP16452286A JPS6322179A JP S6322179 A JPS6322179 A JP S6322179A JP 61164522 A JP61164522 A JP 61164522A JP 16452286 A JP16452286 A JP 16452286A JP S6322179 A JPS6322179 A JP S6322179A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- dried
- carbonate
- bifidus
- sodium bicarbonate
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002253 acid Substances 0.000 title claims abstract description 20
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 claims abstract description 13
- 239000001095 magnesium carbonate Substances 0.000 claims abstract description 13
- 229910000021 magnesium carbonate Inorganic materials 0.000 claims abstract description 13
- 239000003223 protective agent Substances 0.000 claims abstract description 12
- 238000004108 freeze drying Methods 0.000 claims abstract description 6
- 239000007788 liquid Substances 0.000 claims abstract description 5
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 claims description 22
- 241000186000 Bifidobacterium Species 0.000 claims description 18
- 230000001580 bacterial effect Effects 0.000 claims description 12
- 235000017557 sodium bicarbonate Nutrition 0.000 claims description 11
- 229910000030 sodium bicarbonate Inorganic materials 0.000 claims description 11
- 150000004649 carbonic acid derivatives Chemical class 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 241000894006 Bacteria Species 0.000 abstract description 13
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 abstract description 3
- UIIMBOGNXHQVGW-DEQYMQKBSA-M Sodium bicarbonate-14C Chemical compound [Na+].O[14C]([O-])=O UIIMBOGNXHQVGW-DEQYMQKBSA-M 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000000034 method Methods 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 238000007796 conventional method Methods 0.000 abstract 1
- 239000000243 solution Substances 0.000 description 14
- 230000000694 effects Effects 0.000 description 8
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 7
- 239000000843 powder Substances 0.000 description 7
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 6
- CDBYLPFSWZWCQE-UHFFFAOYSA-L Sodium Carbonate Chemical compound [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 description 6
- 230000007423 decrease Effects 0.000 description 5
- 235000020183 skimmed milk Nutrition 0.000 description 5
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 description 4
- 238000007710 freezing Methods 0.000 description 4
- 230000008014 freezing Effects 0.000 description 4
- 235000013923 monosodium glutamate Nutrition 0.000 description 4
- 229940073490 sodium glutamate Drugs 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 239000004201 L-cysteine Substances 0.000 description 3
- 235000013878 L-cysteine Nutrition 0.000 description 3
- 229910000019 calcium carbonate Inorganic materials 0.000 description 3
- 239000007789 gas Substances 0.000 description 3
- 210000004051 gastric juice Anatomy 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- 229910000029 sodium carbonate Inorganic materials 0.000 description 3
- 235000017550 sodium carbonate Nutrition 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 239000001888 Peptone Substances 0.000 description 2
- 108010080698 Peptones Proteins 0.000 description 2
- 229910052782 aluminium Inorganic materials 0.000 description 2
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 2
- VSCWAEJMTAWNJL-UHFFFAOYSA-K aluminium trichloride Chemical compound Cl[Al](Cl)Cl VSCWAEJMTAWNJL-UHFFFAOYSA-K 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 229940069428 antacid Drugs 0.000 description 2
- 239000003159 antacid agent Substances 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 235000019319 peptone Nutrition 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000001488 sodium phosphate Substances 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- ULGYAEQHFNJYML-UHFFFAOYSA-N [AlH3].[Ca] Chemical compound [AlH3].[Ca] ULGYAEQHFNJYML-UHFFFAOYSA-N 0.000 description 1
- YKTSYUJCYHOUJP-UHFFFAOYSA-N [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] Chemical compound [O--].[Al+3].[Al+3].[O-][Si]([O-])([O-])[O-] YKTSYUJCYHOUJP-UHFFFAOYSA-N 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 description 1
- 230000001458 anti-acid effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 229940009291 bifidobacterium longum Drugs 0.000 description 1
- 235000010216 calcium carbonate Nutrition 0.000 description 1
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 description 1
- FUFJGUQYACFECW-UHFFFAOYSA-L calcium hydrogenphosphate Chemical compound [Ca+2].OP([O-])([O-])=O FUFJGUQYACFECW-UHFFFAOYSA-L 0.000 description 1
- 239000000920 calcium hydroxide Substances 0.000 description 1
- 229910001861 calcium hydroxide Inorganic materials 0.000 description 1
- 235000011116 calcium hydroxide Nutrition 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 235000019700 dicalcium phosphate Nutrition 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 235000019800 disodium phosphate Nutrition 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 150000004679 hydroxides Chemical class 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 235000014380 magnesium carbonate Nutrition 0.000 description 1
- 239000000395 magnesium oxide Substances 0.000 description 1
- CPLXHLVBOLITMK-UHFFFAOYSA-N magnesium oxide Inorganic materials [Mg]=O CPLXHLVBOLITMK-UHFFFAOYSA-N 0.000 description 1
- 235000012245 magnesium oxide Nutrition 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- AXZKOIWUVFPNLO-UHFFFAOYSA-N magnesium;oxygen(2-) Chemical compound [O-2].[Mg+2] AXZKOIWUVFPNLO-UHFFFAOYSA-N 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 235000011008 sodium phosphates Nutrition 0.000 description 1
- 229910052938 sodium sulfate Inorganic materials 0.000 description 1
- 235000011152 sodium sulphate Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- RYFMWSXOAZQYPI-UHFFFAOYSA-K trisodium phosphate Chemical compound [Na+].[Na+].[Na+].[O-]P([O-])([O-])=O RYFMWSXOAZQYPI-UHFFFAOYSA-K 0.000 description 1
- 230000035899 viability Effects 0.000 description 1
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、ビフィズス乾燥菌体製造法に関し、生菌数を
長時間の保存においても安定に維持し、しかも酸に対す
る耐性が高い乾燥菌体の効率的な製造法に関するもので
ある。Detailed Description of the Invention (Field of Industrial Application) The present invention relates to a method for producing dried bifidus cells, which maintains the number of viable bacteria stably even during long-term storage and which has high resistance to acids. It concerns an efficient manufacturing method.
（従来の技術） ビフィズス菌は有用な腸内細菌とされている。(Conventional technology) Bifidobacteria are considered to be useful intestinal bacteria.
しかしながら偏性嫌気性菌のため酸素の存在下で死滅し
やすく、又、ｐＨの低い状態においても著しく生菌数が
低下する。However, since it is an obligate anaerobic bacterium, it easily dies in the presence of oxygen, and the number of viable bacteria decreases significantly even in low pH conditions.
このため、ビフィズス菌の乾燥菌体調製においては可及
的に空気に触れないように常に保持し、保持中も脱気、
遮光、低温などの製造条件をきびしく制御しているにも
かかわらず死滅し易く、乾燥後の菌体粉末も一定な生菌
数を長時間にわたって維持することは困難である。For this reason, when preparing dried Bifidobacterium cells, they should always be kept as close to air as possible, and deaerated during the holding process.
Despite strict control of manufacturing conditions such as light shielding and low temperatures, bacteria tend to die, and it is difficult to maintain a constant number of viable bacteria over a long period of time in dried bacterial powder.
一般にビフィズス菌の凍結乾燥には保護剤として脱脂粉
乳や各腫糖類、アミノ酸類などが使用さｈており、これ
らは保護剤としてすぐれた効果を示す。In general, skim milk powder, various saccharides, amino acids, etc. are used as protective agents in the freeze-drying of bifidobacteria, and these exhibit excellent effects as protective agents.
しかし、この場合混合液をがなり低温に保持しても、な
お活発なビフィズス菌の代謝によって産出される酸によ
’）ｐｌ＋の低下が見られるのが普通である。However, in this case, even if the mixed solution is kept at a low temperature, a decrease in pl+ is usually observed due to the acid produced by the metabolism of active bifidobacteria.
このｐＨ低下の程度は生残ビフィズス菌数や温度、時間
により異なるが、大量生産規俣においては作業時間が良
くなるため過度のｐｔ＋低下に達することがある。この
ようなビフィズス菌液を凍結乾燥した場合には凍結１０
客や乾燥障害が者しく凍結収率が低下する。The degree of this pH decrease varies depending on the number of surviving bifidobacteria, temperature, and time, but in mass production, working time becomes shorter, so an excessive decrease in pt+ may be reached. When such a bifidobacteria solution is freeze-dried, the freezing rate is 10
The freezing yield is reduced due to serious drying problems.
また乾燥菌体の保存中における生残性や耐酸性も明らか
に劣るものである。Furthermore, the survivability and acid resistance of dried bacterial cells during storage are clearly inferior.
（発明が解決しようとする問題点）
本発明はビフィズス菌液と保護剤を混合した場合にとフ
イズス菌により産出される酸により保護剤から炭酸〃ス
を発生させて嫌気状態を良好に維持するとともにｐｌ＋
低下を軽減することにより凍結及び乾燥障害を少なくし
、乾燥菌体の生残性改善を見出だす事を最大の問題点と
するものである。(Problems to be Solved by the Invention) The present invention maintains a good anaerobic condition by generating carbon dioxide from the protective agent using the acid produced by the Bifidobacterium when a Bifidobacteria solution and a protective agent are mixed. with pl+
The biggest problem is to find ways to improve the survival of dried microbial cells by reducing the damage caused by freezing and drying.
（問題点を解決するための手段）
本発明は面頂に述べた問題、αを解決することを目的と
するものである。(Means for Solving the Problems) The present invention aims to solve the problem α mentioned above.
本発明者らはこの目的実現のため鋭意研究を重ねていた
ところ、炭酸塩が極く有用なことを発見した。The inventors of the present invention have conducted extensive research to realize this objective, and have discovered that carbonates are extremely useful.
即ち、一般にこのような効果を有する物質としてはナト
リウム、カルシウムアルミニウムなどの金属の酸化物、
水酸化物が考えられる。That is, substances that generally have this effect include metal oxides such as sodium and calcium aluminum;
Possible hydroxides.
本発明者等はビフィズス菌濃厚液を凍結乾燥するに際し
、保護剤として脱脂粉乳、糖類、アミノ酸などを含む混
液中に、各種の塩類を０．１〜１０％添加し、３７℃、
１時間放置後、これらを凍結乾燥した菌体粉末のビフィ
ズス生菌数などを調ヘタ。When freeze-drying a concentrated Bifidobacterium solution, the present inventors added 0.1 to 10% of various salts to a mixed liquid containing skim milk powder, sugars, amino acids, etc. as a protective agent, and heated the mixture at 37°C.
After allowing it to stand for 1 hour, the number of viable bifidus bacteria was determined from the freeze-dried bacterial powder.
使用した塩類は次の通りである。The salts used are as follows.
酢酸ナトリウム、重炭酸ナトリウム、炭酸ナトリウム、
リン酸−ナトリウム、リン酸二ナトリウム、硫酸ナトリ
ウム、水酸化カルシウム、リン酸水素カルシウム、炭酸
カルシウム、硫酸マグネシウム、炭酸マグネシウム、酸
化マグネジツム、硫酸アルミニウム、塩化アルミニウム
、ケイ酸アルミニウム、人工カルルス塩（ｆ５９版 日
本薬局方解説書Ｄ）。Sodium acetate, sodium bicarbonate, sodium carbonate,
Sodium phosphate, disodium phosphate, sodium sulfate, calcium hydroxide, calcium hydrogen phosphate, calcium carbonate, magnesium sulfate, magnesium carbonate, magnesium oxide, aluminum sulfate, aluminum chloride, aluminum silicate, artificial Carlus salt (F59 version) Japanese Pharmacopoeia Commentary D).
その結果、実施例に示すように、胃液の塩酸を中和する
薬剤として一般に広く使用されている炭酸塩のうちで重
炭酸ナトリウム（ｍ曹）と炭酸マグネシウムが特に有効
であることを確認した。As a result, as shown in the Examples, it was confirmed that among the carbonates that are generally widely used as agents for neutralizing hydrochloric acid in gastric juice, sodium bicarbonate (m-carbonate) and magnesium carbonate are particularly effective.
この重炭酸ナトリウムと炭酸マグネシウムが有効である
メカニズムは次のようであると推察される。即ち、これ
らの炭酸塩は冷水に比較的溶は難いが、しかしこのわず
かに溶けた部分は解離の結果として弱アルカリ性を呈す
ると共に炭酸ガスを発生する。このためビフィズス菌の
代謝により産出された酸が中和され、又、炭酸がスによ
り保護されるため、ビフィズス菌の生存に対して有効に
働き、ビフィズス菌乾燥菌体の保存安定性及び耐酸性に
するすぐれたものが得られるように思われる。The mechanism by which sodium bicarbonate and magnesium carbonate are effective is presumed to be as follows. That is, these carbonates are relatively difficult to dissolve in cold water, but this slightly dissolved portion becomes weakly alkaline as a result of dissociation and generates carbon dioxide gas. For this reason, the acid produced by the metabolism of Bifidobacterium is neutralized, and the carbonic acid is protected by SO, which is effective for the survival of Bifidobacterium and improves the storage stability and acid resistance of dried Bifidobacterium cells. It seems like you'll get something great to do with it.
なお、制酸剤としては炭酸カルシウムや炭酸ナトリウム
も広く使用されるものであるが、炭酸カルシウムは第１
表に示すように重炭酸ナトリウム及び炭酸マグネシウム
に比べて溶解度が過小であり、従ってｐＨ低下の阻止が
不充分なために効果が少なく、他方炭酸ナトリウムはこ
れとは逆に、溶解度が過大であり、がっρ１］低下の阻
止限界が高すぎる（ｐｉｔ １１．６）ために効果がほ
とんどないものと推察される。Calcium carbonate and sodium carbonate are also widely used as antacids, but calcium carbonate is the
As shown in the table, the solubility is too low compared to sodium bicarbonate and magnesium carbonate, so it is less effective because it does not sufficiently prevent the pH drop.On the other hand, sodium carbonate, on the other hand, has too much solubility. , ρ1] is too high (pit 11.6), so it is presumed that there is almost no effect.
本発明の好適な実施例を次項に説明する。A preferred embodiment of the invention is described in the following section.
（実施例）
実施例−１
脱脂粉乳７％、酵母エキスＯ０５％、ツイーン８００．
０５％、Ｌ−システィンの０．０５９６などを含む培養
液を加熱滅菌冷却。これにビフイドバクテリワム・ロン
ガムＡ Ｔ ＣＣ１５７０７を接種。培養中１０％Ｎａ
。(Example) Example-1 Skim milk powder 7%, yeast extract O05%, Tween 800.
The culture solution containing 0.05% L-cysteine and 0.0596 L-cysteine was heat sterilized and cooled. This was inoculated with Bifidobacterium longum AT CC15707. 10% Na during culture
.
ＣＯ３を添加してｐｉｔ６．０以上に維持しＣ○２ガス
、Ｎ２ガスを通じながら３７℃、８時間の嫌気培養を行
い、培養終了後、遠心分離により濃厚菌液をＩＴＩ製し
た。Anaerobic culture was carried out at 37° C. for 8 hours while adding CO3 to maintain the pit at 6.0 or higher and passing C2 gas and N2 gas. After completion of the culture, a concentrated bacterial solution was prepared by centrifugation using ITI.
これに脱脂粉乳２５％、デンプン分解物１０％、グルタ
ミン酸ナトリウム１％含む保護剤溶液をｐＨ７，０に１
４整後、重炭酸ナトリウム、若しくは炭酸マグネシウム
又はその両者を混合して加えよく分１牧させた。To this, add a protectant solution containing 25% skim milk powder, 10% starch decomposition product, and 1% sodium glutamate to a pH of 7.0.
After 4 hours, sodium bicarbonate or magnesium carbonate, or a mixture of both, was added and stirred well.
重炭酸ナトリウム、炭酸マグネシウムの保護剤溶液への
添加量は０．５％以下では効果がなく１０％以上ではｐ
ｉｔの上昇が大きい。このため、経済性と呈味性を：ｉ
ｆａ！ して添加量１％〜７．５％を選んだ。If the amount of sodium bicarbonate or magnesium carbonate added to the protectant solution is less than 0.5%, it will not be effective, and if it is more than 10%, it will be harmful.
The increase in IT is large. For this reason, economic efficiency and taste: i
fa! The addition amount was selected from 1% to 7.5%.
このようにして調整した保護剤溶液に前記濃ｊ７菌液を
加え混合した。The concentrated j7 bacteria solution was added to the protective agent solution thus prepared and mixed.
ついでこれらをそれぞれ半量に分け、一方を直ちに凍結
乾燥、他方を３７℃１時間放置後、凍結乾燥を行った。Next, each of these was divided into halves, and one portion was immediately freeze-dried, and the other portion was left to stand at 37° C. for 1 hour, and then freeze-dried.
なお、重炭酸ナトリウム、炭酸マグネシウムを加えない
ものを対照として制酸作用、凍結乾燥収率、乾燥菌体の
保存安定性、耐酸性について比較した。In addition, the antacid effect, freeze-drying yield, storage stability of dried bacterial cells, and acid resistance were compared using a control sample to which no sodium bicarbonate or magnesium carbonate was added.
その結果、第２表、第３表に示すように明らかに重炭酸
ナトリワム、炭酸マグネシウムの効果が認められた。As a result, as shown in Tables 2 and 3, the effects of sodium bicarbonate and magnesium carbonate were clearly observed.
第３表；耐酸性（実施例−１）車
京耐酸性試験法
人工胃液（＃母エキス１．２５ｇ、ペプトン２．５ｇ、
乳糖０．５ｇ、ツイーン８０ Ｑ、５ｇ、Ｌシ入ティン
０．０５．、塩化ナトリウム１．０８を蒸留水５００ｍ
１で溶解後滅菌）に乾燥菌体ヲ入７ｔ、Ｉ Ｎ ＨＯ２
”ｌ’ｐＨ３，０１：ＷＩ４９１Ｊｚ。Table 3: Acid resistance (Example-1) Chejing acid resistance test method Artificial gastric juice (#Mother extract 1.25g, peptone 2.5g,
Lactose 0.5g, Tween 80 Q, 5g, L Shitin 0.05. , sodium chloride 1.08 in distilled water 500m
7 tons of dried bacterial cells (sterilized after dissolving in step 1), IN HO2
"l'pH3,01: WI491Jz.
これを３７℃１時間、３ＩＬ！間放置し、直ちにｐＨ６
，０に？Ｉ４整し、生菌数を測定した。Cook this for 1 hour at 37℃ for 3IL! Leave it for a while and immediately adjust the pH to 6.
, to 0? I4 was adjusted and the number of viable bacteria was measured.
実施例−２
実施例−１において使用したと同じ組成の液体培地で濃
厚菌液をＩＩ！！ｓ！した。Example-2 A concentrated bacterial solution was prepared using a liquid medium with the same composition as used in Example-1! ! s! did.
これに脱脂粉乳１２％、砂糖５％、ゼラチン１．２％、
グルタミン酸ナトリウム１．０％を含む保護剤溶液のｐ
Ｈを７．０に調整後、重炭酸ナトリウム、炭酸マグネシ
ウムをそれぞれ加えた後よく分散させた。Add to this 12% skim milk powder, 5% sugar, 1.2% gelatin,
p of protectant solution containing 1.0% sodium glutamate
After adjusting H to 7.0, sodium bicarbonate and magnesium carbonate were added and well dispersed.
これに前記濃厚菌液を加え、よく混合、実施例−１と同
様な方法で重炭酸ナトリウム、炭酸マグネシウムの効果
を調べた。The above-mentioned concentrated bacterial solution was added and mixed well, and the effects of sodium bicarbonate and magnesium carbonate were investigated in the same manner as in Example-1.
その結果、＃Ｓ４表、第５表に示すように明らかに重炭
酸ナトリウム、炭酸マグネシウムの効果が認められた。As a result, as shown in Tables #S4 and 5, the effects of sodium bicarbonate and magnesium carbonate were clearly observed.
ｔｐＪＳｌ＆；耐酸性（実施例−２）本本耐酸性試験法
人工胃液（酵母エキス１．２５ｇ、ペプトン２．５ｇ、
乳糖０．５ｇ、ツイーン８００，５ｇ。tpJSl&;Acid resistance (Example-2) This acid resistance test method Artificial gastric juice (yeast extract 1.25g, peptone 2.5g,
Lactose 0.5g, Tween 800.5g.
Ｌシスティン０．０５ｇ、塩化ナトリウム１．Ｏｇを蒸
留水５００−で溶解ａ滅菌）に乾燥菌体を入れ、ＩＮＨ
（Ｊ’でｐｌ＋ ３．０に、ｉ！ｌ整した。L-cysteine 0.05g, sodium chloride 1. Dissolve Og in distilled water (sterilized) and add the dried bacterial cells to INH.
(I adjusted i!l to pl+ 3.0 with J'.
これを３７℃１時間、３時間放置し、直ちにｐ＋＋ ６
．０に調整し、生菌数を測定した。This was left at 37°C for 1 hour and 3 hours, and immediately p++ 6
．． The number of viable bacteria was measured.
（本発明の作用及び効果）
本発明は前記した説明により明らかにしたように、一般
にビフィズス菌の凍結乾燥に供用される保護剤の、ビフ
ィズス菌から産出される酸によ’）ｐｌ＋の低下が見ら
れ、嫌気状態を不良にして凍結障害や乾燥障害を米たし
、かつ乾燥菌体の保存中における生残性 耐酸性を劣ら
せるのを、炭酸塩の保護剤への添加により、生残性と耐
酸性にすぐれたビフィズス菌乾燥体を製造できるもので
、極く有用な発明をなすものである。(Actions and Effects of the Present Invention) As clarified from the above explanation, the present invention provides a protective agent that is generally used for freeze-drying Bifidobacterium, and that the acid produced by Bifidobacterium causes a decrease in pl+. The addition of carbonate to the protective agent improves the survival of dried microorganisms, which reduces freezing and drying damage due to poor anaerobic conditions and reduces the survival and acid resistance of dried bacterial cells during storage. This invention makes it possible to produce a dried bifidobacterium with excellent properties and acid resistance, making it an extremely useful invention.
出願人 新日本化学工業 株式会社
手続補正書
１．事件の表示 特願昭６１−１６４５２２２、発明の
名称 生残性及びａｔ酸性ビフィズス菌乾燥菌体製造法
３、補正をする者
事件との関係 出願人
名 称 新日本化学工業株式会社
４、代理人
６、補正により増加する発明の数
７、補正の対象 明細書
補 正 の 内 穿
１、明細書第４頁第１３行に「カルシウムアルミニウム
」とあるを
「カルシウム、アルミニウム」
と訂正する。Applicant Shin Nippon Chemical Co., Ltd. Procedural Amendment 1. Indication of the case: Japanese Patent Application No. 1645222/1983, title of the invention: Viability and at-acid bifidobacterium dried bacterial cell production method 3, person making the amendment Relationship with the case: Applicant name Title: Shin Nippon Chemical Co., Ltd. 4, Agent 6 , the number of inventions increased by the amendment 7, the subject of the amendment, paragraph 1 of the amendment to the specification, the phrase ``calcium aluminum'' in line 13 of page 4 of the specification is corrected to ``calcium, aluminum.''
２、明細書ｅｊＳ６頁第１０〜１１行に「耐酸性１こす
るすぐれたもの」とあるを
［耐酸性にすぐれたちの」
と訂正する。2. In the specification ejS, page 6, lines 10-11, the phrase "excellent acid resistance 1 rub" is corrected to "excellent acid resistance."
３、明細書第８頁第７〜８行に「グルタミン酸ナトリウ
ム１％含む保護剤溶液」とあるを「グルタミン酸ナトリ
ウム１９６を含む保護剤溶液」と訂正する。3. On page 8, lines 7-8 of the specification, the phrase "protectant solution containing 1% sodium glutamate" is corrected to "protectant solution containing 196 sodium glutamate."
以 上that's all
Claims (1)
保護剤中に重炭酸ナトリウム、炭酸マグネシウム等の炭
酸塩を加え、凍結乾燥することを特徴とする生残性と耐
酸性のすぐれたビフィズス乾燥菌体製造法。Bifidobacteria drying with excellent survivability and acid resistance is achieved by adding carbonates such as sodium bicarbonate and magnesium carbonate to a known protective agent commonly used for concentrated bifidobacteria culture liquids and freeze-drying the mixture. Bacterial cell production method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61164522A JPS6322179A (en) | 1986-07-11 | 1986-07-11 | Production of survivable and acid-resistant dried bifidus cell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61164522A JPS6322179A (en) | 1986-07-11 | 1986-07-11 | Production of survivable and acid-resistant dried bifidus cell |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6322179A true JPS6322179A (en) | 1988-01-29 |
| JPH0465677B2 JPH0465677B2 (en) | 1992-10-20 |
Family
ID=15794760
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61164522A Granted JPS6322179A (en) | 1986-07-11 | 1986-07-11 | Production of survivable and acid-resistant dried bifidus cell |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6322179A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2075807A1 (en) * | 1994-01-05 | 1995-10-01 | Ind Lacteas Talavera S A | Improved system for fermenting milk products to obtain bifidus |
| WO2009157073A1 (en) * | 2008-06-26 | 2009-12-30 | 信和薬品株式会社 | Nano-level lactic acid bacterium |
| RU2560425C2 (en) * | 2008-08-28 | 2015-08-20 | Кр. Хансен А/С | Stabiliser of colour in bacterial composition |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7552781B2 (en) | 2004-10-20 | 2009-06-30 | Black & Decker Inc. | Power tool anti-kickback system with rotational rate sensor |
| US9475180B2 (en) | 2010-01-07 | 2016-10-25 | Black & Decker Inc. | Power tool having rotary input control |
| US8418778B2 (en) | 2010-01-07 | 2013-04-16 | Black & Decker Inc. | Power screwdriver having rotary input control |
| US9266178B2 (en) | 2010-01-07 | 2016-02-23 | Black & Decker Inc. | Power tool having rotary input control |
-
1986
- 1986-07-11 JP JP61164522A patent/JPS6322179A/en active Granted
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2075807A1 (en) * | 1994-01-05 | 1995-10-01 | Ind Lacteas Talavera S A | Improved system for fermenting milk products to obtain bifidus |
| WO2009157073A1 (en) * | 2008-06-26 | 2009-12-30 | 信和薬品株式会社 | Nano-level lactic acid bacterium |
| JPWO2009157073A1 (en) * | 2008-06-26 | 2011-12-01 | 信和薬品株式会社 | Nano-type lactic acid bacteria |
| JP5257363B2 (en) * | 2008-06-26 | 2013-08-07 | 信和薬品株式会社 | Method for producing cells of nano-type lactic acid bacteria |
| RU2560425C2 (en) * | 2008-08-28 | 2015-08-20 | Кр. Хансен А/С | Stabiliser of colour in bacterial composition |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0465677B2 (en) | 1992-10-20 |
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