JPS5811193B2 - Method for producing bacterial cells - Google Patents
Method for producing bacterial cellsInfo
- Publication number
- JPS5811193B2 JPS5811193B2 JP1398181A JP1398181A JPS5811193B2 JP S5811193 B2 JPS5811193 B2 JP S5811193B2 JP 1398181 A JP1398181 A JP 1398181A JP 1398181 A JP1398181 A JP 1398181A JP S5811193 B2 JPS5811193 B2 JP S5811193B2
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- JP
- Japan
- Prior art keywords
- bacterial cells
- culture solution
- culture
- dried
- methylamine
- Prior art date
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Description
【発明の詳細な説明】
本発明は、メチルアミン類資化性細菌によって細菌菌体
を製造する方法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing bacterial cells using methylamine-assimilating bacteria.
細菌菌体は、飼料もしくは食品または医薬用原料もしく
は工業用原料として多量に使用されている。Bacterial cells are used in large quantities as feed, food, pharmaceutical raw materials, or industrial raw materials.
従来、微生物菌体の製造原料としては、糖蜜、亜硫酸パ
ルプ廃液およびｎ−パラフィンなどが使用されてきた。Conventionally, molasses, sulfite pulp waste liquid, n-paraffin, and the like have been used as raw materials for producing microbial cells.
しかしながら、これらの物質は、醗酵原料としては供給
量および価格の点で不安定であり問題がある。However, these substances are unstable and problematic as fermentation raw materials in terms of supply amount and price.
さらに、ｎ−パラフィンは水に難溶性乃至不溶性である
ことおよび微生物菌体の生産に多量の酸素が必要とされ
る点などで培養上問題が多い。Furthermore, n-paraffin poses many problems in terms of cultivation because it is sparingly soluble or insoluble in water and requires a large amount of oxygen for the production of microbial cells.
一方、メチルアミン、ジメチルアミン、およびトリメチ
ルアミンなどのメチルアミン類は化学工業により大量に
得られ、しかも水溶性が大であり、かつ分子内に炭素原
子とともに窒素原子を含有し、炭素源であるとともに窒
素源であることなどから、醗酵原料として好ましい物質
である。On the other hand, methylamines such as methylamine, dimethylamine, and trimethylamine are obtained in large quantities through the chemical industry, are highly water-soluble, contain nitrogen atoms as well as carbon atoms in their molecules, and serve as a carbon source. It is a preferred material as a fermentation raw material because it is a nitrogen source.
本発明者は、他の炭素源の不存在下でもメチルアミン、
ジメチルアミンおよびトリメチルアミンの少なくとも１
種のみを炭素源として強力に資化する細菌の検索を行な
ったところ、アクロモバクタ−メタノロヒラ（Ａｃｈｒ
ｏｍｏｂａｃｔｅｒ ｍｅｔｈａｎｏｌｏ−ｐｈｉｌａ
）ＡＴＣＣ２１２７５、メチロバクテリウム オルガ
ノヒラム（Ｍｅｔｈｙｌｏｂａｃｔｅｒｉｕｍ ｏｒｇ
ａｎｏｐ−ｈｉｌｕｍ）ＸＸ ＡＴＣＣ２７８８６Ｉ
ｎｔ−Ｊ−８ｙｓｔ。The inventor has demonstrated that methylamine, even in the absence of other carbon sources,
at least one of dimethylamine and trimethylamine
When searching for bacteria that strongly assimilate seeds using only seeds as a carbon source, we found that Achromobacter methanorohira (Achr.
omobacter methanolo-phila
) ATCC21275, Methylobacterium org.
anop-hilum)XX ATCC27886I
nt-J-8yst.
Ｂａｃｔ−２６：２２６−２２９（１９７６））、ミク
ロチクルス アキュアテイクス（Ｍｉｃｒｏｃｙｃｌｕ
ｓａｑｕａｔｉｃｕｓ ） ＡＴＣＣ２７０６８、ミク
ロチクルス エブルネウス（Ｍｉｃｒｏｃｙｃｌｕｓ
ｅｂｒｕｎｅｕｓ ） ＡＴＣＣ２１３７３、ミクロチ
クルス ポリモルファム（Ｍｉ−ｃｒｏｃｙｃｌｕｓ
ｐｏｌｙｍｏｒｐｈｕｍ ） ＮＣＩ Ｂ １０５１
６、ミクロチクルス メタノリカ（Ｍｉｃｒｏｃｙｃｌ
ｕｓ ｍｅｔｈ−ａｎｏｌｉｃａ ） Ｃ−４２微工研
菌寄第４４１８号、パラコツカス デニトリフィカンス
（Ｐａｒａｃｏｃｃｕｓ ｄ−ｅｎｉｔｒｉｆｉｃａｎ
ｓ ） ＡＴＣＣ１３５４３およびチオバチルス エス
ピー（’ｒｈｉｏｂａｃｉｌｌｕｓ ５ｐ−）Ａ２
ＡＴＣＣ２５３６４Ｊ−Ｂａｃｔ、１００ ： ４８７
−４９７（１９６９））などがそれぞれメチルアミン類
を資化して生育、増殖することを見出し、本発明を完成
した。Bact-26:226-229 (1976)), Microcycle
saquaticus ATCC27068, Microcyclus ebruneus
ebruneus) ATCC21373, Microticulus polymorphum (Mi-crocyclus
polymorphum) NCI B 1051
6. Microcyclus methanolica
us meth-anolica) C-42 Microtechnical Laboratory No. 4418, Paracoccus d-enitrifican
s) ATCC13543 and Thiobacillus sp. ('rhiobacillus 5p-) A2
ATCC25364J-Bact, 100: 487
-497 (1969)) and the like grow and proliferate by assimilating methylamines, and completed the present invention.
なお、これらの菌株はいずれも少なくともメチルアミン
を資化するが、これらの中にはさらにジメチルアミンお
よび／またはトリメチルアミンをも資化するものもある
。All of these strains assimilate at least methylamine, but some of these strains also assimilate dimethylamine and/or trimethylamine.
なお、前記において、”ＡＴＣＣ”は［ジ アメリカン
タイプ カルチャー コレクション（ＴｈｅＡｍｅｒ
ｉｃａｎ Ｔｙｐｅ Ｃｕ１ｔｕｒｅ Ｃｏ１１ｅｃｔ
ｉｏｎｔ １２３０１Ｐａｒｋｌａｗｎ Ｄｒｉｖｅ
−Ｒｏｃｋｖｉｌｌｅ３Ｍｄ −２０８５２、Ｕ、Ｓ、
Ａ）ｊの、また”ＮＣＩＢ”は［ナショナル コレクシ
ョン オブ インダストリアルバクテリア （Ｎａｔｉ
ｏｎａｌ Ｃｏ１１ｅｃｔｉｏｎ ｏｆ Ｉｎｄｕｓｔ
ｒｉ−ａｌ Ｂａｃｔｅｒｉａ、 Ａｂｅｒｄｅｅｎ、
５ｃｏｔｌａｎｄ）ｊの、また、”微工研″は、「工
業技術院微生物工業技術研究所」の略称であり、細菌が
これらの機関にそれぞれ保存されていることを示す（以
下同様）。In addition, in the above, "ATCC" refers to [The American Type Culture Collection].
ican Type Culture Co11ect
iont 12301Parklawn Drive
-Rockville3Md-20852, U, S,
A) j, and “NCIB” are from the National Collection of Industrial Bacteria (Nati
onal Co11ection of Industry
ri-al Bacteria, Aberdeen,
5cotland)j, and "Feikoken" is an abbreviation for "National Institute of Microbial Technology, Agency of Industrial Science and Technology", indicating that bacteria are stored in these institutions (the same applies hereinafter).
すなわち本発明は、アクロモバクタ−属、メチロバクテ
リウム属、ミクロチクルス属、パラコツカス属またはチ
オバチルス属に属し、メチルアミン類を資化しうる細菌
を、メチルアミン類を主たる炭素源とする培地に培養し
該培養液から細菌の菌体を分離することを特徴とする細
菌菌体の製造方法である。That is, the present invention involves culturing bacteria that belong to the genus Achromobacter, Methylobacterium, Microticulus, Paracoccus, or Thiobacillus and can assimilate methylamines in a medium containing methylamines as the main carbon source. This is a method for producing bacterial cells, which is characterized by separating bacterial cells from the culture solution.
本発明での細菌は、アクロモバクタ−属、メチロバクテ
リウム属、ミクロチクルス属、バラコツカス属またはチ
オバチルス属に属し、メチルアミン類を資化しうる細菌
であればよく、代表例として前記の細菌が挙げられる。The bacteria used in the present invention may belong to the genus Achromobacter, Methylobacterium, Microticulus, Balacoccus, or Thiobacillus and can assimilate methylamines, and the above-mentioned bacteria are representative examples. It will be done.
メチルアミン類の資化性の有無および強さを検討するた
めに次のような液体培地を用いた。The following liquid medium was used to examine the assimilation ability and strength of methylamines.
すなわち、（ＮＨ４） ２ Ｓ ０４３ ｆｉ、ＫＨ２
ＰＯ４１，４ｇＮａ ２ＨＰＯ４２，１＆、 Ｍｇ５Ｏ
，・７Ｈ２００，２ｇ、ＣａＣａＣｌ２２Ｈ２Ｏ３０、
ＦｅＣ６Ｈ５０７・ＸＨ，，０３０ｍ９、ＭｎＣ１！２
４Ｈ２０５ｍｌ？、Ｚｎ５Ｏ，’ ７Ｈ２０５ｗｕｉｌ
、 ＣｕＳＯ４・５Ｈ２００、５１１’Ｑ、メチルアミ
ン塩酸塩、ジメチルアミン塩酸塩またはトリメチルアミ
ン塩酸塩 １０ｇおよび下記の組成を有するビタミン混
合液 ０．１ ｍｌを純水 １１に溶解し、ｐＨを６５
に調整して１ｋｇ／ｃｍ２Ｇで２０分間殺菌した液体培
地である。That is, (NH4) 2 S 043 fi, KH2
PO41,4gNa 2HPO42,1&, Mg5O
,・7H200,2g, CaCaCl22H2O30,
FeC6H507・XH, 030m9, MnC1!2
4H205ml? ,Zn5O,' 7H205wuil
, CuSO4・5H200, 511'Q, 10 g of methylamine hydrochloride, dimethylamine hydrochloride or trimethylamine hydrochloride and 0.1 ml of a vitamin mixture having the following composition were dissolved in pure water 11, and the pH was adjusted to 65.
This is a liquid medium that has been sterilized at 1 kg/cm2G for 20 minutes.
ビタミン混合液の組成は、ビオチン ２０ｇｇパントテ
ン酸カルシウム ４■、葉酸 ２０ｇｇイノシトール
２０■、ニコチン酸 ４ｍｇ、ピリドキシン塩酸塩 ４
ｍｇ、チアミン塩酸塩、 ４ｍｇパラ−アミノ安息香酸
２ｍｇ、リボフラビン ２ｍｇ、純水あるいは蒸留水
１ ｍｌである。The composition of the vitamin mixture is: biotin 20gg calcium pantothenate 4■, folic acid 20gg inositol
20■, Nicotinic acid 4mg, Pyridoxine hydrochloride 4
mg, thiamine hydrochloride, 4 mg, para-aminobenzoic acid 2 mg, riboflavin 2 mg, and pure or distilled water 1 ml.
本細菌の培養に使用する培地は、メチルアミン類を主た
る炭素源として含有していることを要しさらに窒素源お
よび無機物などの適量を含有する培地ならば合成培地又
は天然培地のいずれでもよい。The medium used for culturing this bacterium must contain methylamines as a main carbon source, and may be either a synthetic medium or a natural medium as long as it further contains appropriate amounts of a nitrogen source and inorganic substances.
メチルアミン類とはメチルアミン、ジメチルアミンおよ
びトリメチルアミンであって、これらのうち使用する細
菌が資化しうるアミンを使用するこれらのメチルアミン
類としてメチルアミン、ジメチルアミンおよびトリメチ
ルアミンそれ自体のほかにこれらの塩も使用しうる。Methylamines are methylamine, dimethylamine, and trimethylamine, and these methylamines include amines that can be assimilated by the bacteria used. Salt may also be used.
これらの塩としては塩酸塩、硝酸塩および硫酸塩などの
無機酸塩ならびにぎ酸塩および酢酸塩などの有機酸塩を
使用しうる。As these salts, inorganic acid salts such as hydrochlorides, nitrates and sulfates and organic acid salts such as formates and acetates can be used.
これらの中で、実用上メチルアミン塩酸塩、ジメチルア
ミン塩酸塩およびトリメチルアミン塩酸塩などの塩酸塩
を用いることが好ましい。Among these, it is practically preferable to use hydrochlorides such as methylamine hydrochloride, dimethylamine hydrochloride, and trimethylamine hydrochloride.
またこれらの混合物も使用しうる。Mixtures of these may also be used.
培養液中のメチルアミン類の濃度はメチルアミン、ジメ
チルアミンまたはトリメチルアミンとして１重量％以下
が好ましく、菌の生育および増殖の良好さからは、０．
５重量％以下であることが特に好ましい。The concentration of methylamines in the culture solution is preferably 1% by weight or less as methylamine, dimethylamine or trimethylamine, and from the viewpoint of good growth and multiplication of bacteria, the concentration is 0.5% by weight or less.
It is particularly preferable that the amount is 5% by weight or less.
窒素源としては、たとえばアンモニウム塩および硝酸塩
などの無機窒素化合物または、たとえば尿素、コーン・
ステイープ・リカー、カゼイン、ペプトン、酵母エキス
および肉エキスなどの有機窒素含有物が用いられる。As nitrogen sources, inorganic nitrogen compounds such as ammonium salts and nitrates or e.g. urea, corn
Organic nitrogen-containing substances are used, such as staple liquor, casein, peptone, yeast extract and meat extract.
その他、無機化合物として、たとえばカルシウム塩、マ
グネシウム塩、カリウム塩、ナトリウム塩、リン酸塩、
マンガン塩、亜鉛塩、鉄塩、銅塩、モリブデン塩および
コバルト塩ならびにホウ素化合物およびヨウ素化合物な
どが用いられる。Other inorganic compounds include calcium salts, magnesium salts, potassium salts, sodium salts, phosphates,
Manganese salts, zinc salts, iron salts, copper salts, molybdenum salts and cobalt salts, boron compounds and iodine compounds, and the like are used.
培養条件は、温度２０〜４０°Ｃ１好ましくは２５〜３
７℃、およびｐＨは５〜９、好ましくは６〜８である。Culture conditions include temperature 20-40°C, preferably 25-30°C.
7°C, and the pH is 5-9, preferably 6-8.
このような条件で好気的に培養を行なう。これらの条件
をはずれて培養した場合には、本細菌の増殖は悪くなる
。Culture is carried out aerobically under these conditions. If cultured outside these conditions, the growth of this bacterium will be impaired.
また、培養液中の溶存酸素濃度には特に制限はないが、
実用上、通常は０．５〜２０ＰＰｍが好ましい。In addition, there is no particular limit to the dissolved oxygen concentration in the culture solution, but
Practically speaking, 0.5 to 20 PPm is usually preferable.
そのために通気量を調節したり、撹拌したり、又、培養
槽内の圧力を高めるなどの手段が採用される。For this purpose, measures such as adjusting the amount of ventilation, stirring, and increasing the pressure inside the culture tank are adopted.
また、培養方式は、回分培養または連続培養のいずれで
もよい。Furthermore, the culture method may be either batch culture or continuous culture.
窒素源としてアンモニウム塩を利用する場合は、培養期
間中にアンモニアが菌体生成のため消費されて培養液の
ｐＨが低下する。When ammonium salt is used as a nitrogen source, ammonia is consumed during the culture period due to bacterial cell production, resulting in a decrease in the pH of the culture solution.
従って、培養液のｐＨを一定に保つために、アンモニア
、カセイカリ、又は、カセイソーダなどのアルカリを添
加する。Therefore, in order to keep the pH of the culture solution constant, an alkali such as ammonia, caustic potash, or caustic soda is added.
これらのうち、アンモニアを添加することが好ましい。Among these, it is preferable to add ammonia.
このようにして細菌を培養した後、菌体を培養液より分
離する。After culturing the bacteria in this manner, the bacterial bodies are separated from the culture solution.
分離の手段としては、培養液そのままを遠心分離すると
の手段、培養液中に本細菌よりも大きい他の微生物の細
胞をろ過励剤として加えたり、あるいはプレコートする
ことにより培養液から菌体をろ過分離するとの手段、培
養液中に種々の凝集剤を加え、菌体を凝集させ、これを
ろ過あるいは遠心分離により培養液から菌体を分離する
との手段、培養液のｐＨを５以下にすることにより、も
しくはｐＨを５以下にさらに５０〜１００℃で加熱する
ことにより菌体を凝集させ、これをろ過あるいは遠心分
離により菌体を分離するとの手段などを適用しうる。Separation methods include centrifuging the culture solution as it is, adding cells of other microorganisms larger than the present bacteria to the culture solution as a filtration promoter, or filtering the bacterial cells from the culture solution by pre-coating. Means for separating, adding various flocculants to the culture solution to agglutinate the bacteria, and separating the bacteria from the culture solution by filtration or centrifugation, and adjusting the pH of the culture solution to 5 or less. Alternatively, the microbial cells may be aggregated by further heating at 50 to 100° C. to a pH of 5 or less, and the microbial cells may be separated by filtration or centrifugation.
分離された菌体を、必要ならば各種の有機溶媒あるいは
水で洗浄し湿潤菌体を得る。If necessary, the separated bacterial cells are washed with various organic solvents or water to obtain wet bacterial cells.
湿潤菌体は乾燥され、そのままあるいは、さらに種々の
処理がほどこされた後、飼料などとして利用される。The wet bacterial cells are dried and used as feed or the like after being subjected to various treatments.
また、得られた菌体から、さらに核酸、ビタミン類、補
酵素、蛋白質または脂質などの菌体含有物質が単独の物
質として、またはこれら相互の混合物として抽出され、
飼料、食品、医薬品および工業原料などに使用される。Further, from the obtained bacterial cells, substances contained in the bacterial cells such as nucleic acids, vitamins, coenzymes, proteins, or lipids are extracted as individual substances or as a mixture of these substances,
Used in feed, food, medicine, and industrial raw materials.
本発明によって、工業生産により、容易に入手しうる物
質であるメチルアミン類を原料として使用することがで
き、しかも高収率で蛋白含量の高い菌体を得ることが出
来、かつ原料物質のメチルアミン類は水溶性であるため
培養が容易であり、簡単な後処理で清浄な菌体を得るこ
とが出来、工業上極めて有利な方法である。According to the present invention, methylamines, which are easily available substances, can be used as a raw material through industrial production, and bacterial cells with high protein content can be obtained with high yield, and methyl amines as a raw material can be obtained. Since amines are water-soluble, they are easy to culture, and clean microbial cells can be obtained with simple post-treatment, making this an extremely advantageous method industrially.
実施例によって、さらに詳しく説明する。This will be explained in more detail with reference to examples.
実施例 １
純水１１あたり（ＮＨ４）２ＳＯ４３ｇ、ＫＨ２ＰＯ４
１，４ｇ、Ｎａ ２ＨＰ０４２．１．９ 、、、ＭｎＣ
ｌ２−４Ｈ２０５ｍｇ、ＺｎＳＯ４’ ７Ｈ２０５ｍ？
、ＣｕＳＯ４’ ５Ｈ２００，５ｍ９、ＭｇＳＯ４・７
Ｈ２００，２ｇ、ＣａＣｌ２・２２Ｈ２Ｏ３０？、Ｆｅ
Ｃ６Ｈ５０７・ＮＨ２０３０ｍ９およびメチルアミン塩
酸塩 １０ｇを溶解し、さらにビタミン混合液 ０．１
ｍｌを加え、ｐＨが６．５に調整された液 ５００ｍ
１を１１容ミニ・ジャーに入れ１２０°Ｃで２０分間殺
菌した後、メタノール ５ｇを無菌的に添加し、これを
培地とした。Example 1 (NH4)2SO43g, KH2PO4 per 11 parts of pure water
1.4g, Na 2HP042.1.9 , , MnC
l2-4H205mg, ZnSO4' 7H205m?
, CuSO4' 5H200,5m9, MgSO4・7
H200, 2g, CaCl2・22H2O30? , Fe
Dissolve 10g of C6H507/NH2030m9 and methylamine hydrochloride, and add 0.1 g of vitamin mixture.
ml and adjusted the pH to 6.5 500ml
1 was placed in an 11 volume mini jar and sterilized at 120°C for 20 minutes, then 5 g of methanol was added aseptically and this was used as a medium.
これに上記と同様な培地を用いて３７°Ｃで３日間前培
養されたアクロモバクタ−メタノロヒラＡＴＣＣ２１２
７５の菌体を含む前培養液を上記の培地に３容量％接種
し、培養期間中の培養液のｐＨが６．５に維持されるよ
うにアンモニア水を添加しながら、３０℃で通気撹拌培
養を行なった。This was precultured with Achromobacter methanorohila ATCC212 at 37°C for 3 days using the same medium as above.
A preculture solution containing 75 bacterial cells was inoculated at 3% by volume into the above medium, and aqueous ammonia was added to maintain the pH of the culture solution at 6.5 during the culture period, while stirring at 30°C with aeration. Culture was performed.
２日間培養後、培養液中のメチルアミン濃度は０．００
１ｗｔ％ 以下となった。After 2 days of culture, the methylamine concentration in the culture solution was 0.00.
It became less than 1wt%.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５°Ｃで２４時間乾燥して培養液１１あたり、１
．７ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105°C for 24 hours to give 1
．． 7 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約８３％であった。The crude protein content of the obtained bacterial cells was approximately 83%.
実施例 ２
実施例１と同様な培地、培養条件で、メチロバクテリウ
ム オルガノヒラム ＸＸ ＡＴＣＣ２７８８６を培
養した。Example 2 Methylobacterium organohylum XX ATCC27886 was cultured using the same medium and culture conditions as in Example 1.
３日間培養した後、培養液中のメチルアミン濃度は、０
．００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５℃で２４時間乾燥して、培養液１１あたり１．
６ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105° C. for 24 hours.
6 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約８０％であった。The crude protein content of the obtained bacterial cells was approximately 80%.
実施例 ３
実施例１と同様な培地、培養条件で、ミクロチクルス
アキュアテイクス ＡＴＣＣ２７０６８を培養した。Example 3 Using the same medium and culture conditions as in Example 1, microticuli
Acuretakes ATCC27068 was cultured.
３日間培養した後、培養液中のメチルアミン濃１度は０
．０００１ ｗｔ％以下となった。After culturing for 3 days, the concentration of methylamine in the culture solution was 0.
．． 0001 wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５℃で２４時間乾燥して培養液１１あたり１．５
ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and collect the bacterial cells, which were dried at 105°C for 24 hours to give a
g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は、約７９％であった。The crude protein content of the obtained bacterial cells was about 79%.
実施例 ４
実施例１と同様な培地、培養条件でミクロチクルス エ
ブルネウス ＡＴＣＣ２１３７３を培養した。Example 4 Microticulus ebruneus ATCC21373 was cultured using the same medium and culture conditions as in Example 1.
３日間培養した後、培養液中のメチルアミン濃度は、０
．００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５°Ｃで２４時間乾燥して、培養液１１あたり１
．５ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105°C for 24 hours.
．． 5 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は、約８０％であった。The crude protein content of the obtained bacterial cells was approximately 80%.
実施例 ５
実施例１と同様な培地、培養条件でミクロチクルス ポ
リモルファム ＮＣＩＢ １０５１６を培養した。Example 5 Microticulus polymorphum NCIB 10516 was cultured in the same medium and culture conditions as in Example 1.
３日間培養した後、培養液中のメチルアミン濃度は、０
．００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して、菌体を分離回収し、この菌
体を１０５℃で２４時間乾燥して培養液１１あたり１．
４ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105° C. for 24 hours to yield 1.
4 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約８１％であった。The crude protein content of the obtained bacterial cells was approximately 81%.
実施例 ６
実施例１と同様な培地、培養条件で、ミクロチクルス
メタノリカ 微工研菌寄第４４１８号の培養を行なった
。Example 6 Using the same medium and culture conditions as in Example 1, microticuli
Methanolica Kaikoken Bacillus No. 4418 was cultured.
３日間培養した後、培養液中のメチルアミン濃度は０．
００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
001wt% or less.
この培養液を遠心分離して、菌体を分離回収し、この菌
体を１０５℃で２４時間乾燥して、培養液１１あたり１
．５ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were then dried at 105°C for 24 hours.
．． 5 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は、約８０％であった。The crude protein content of the obtained bacterial cells was approximately 80%.
実施例 ７
実施例１と同様な培地、培養条件で、パラコツカス デ
ニトリフィカンス ＡＴＣＣ１，３５４３を培養した。Example 7 Paracoccus denitrificans ATCC 1,3543 was cultured using the same medium and culture conditions as in Example 1.
３日間培養した後、培養液中のメチルアミン濃度は、０
．００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５℃で２４時間乾燥して、培養液１１あたり１．
４ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105° C. for 24 hours.
4 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約７７％であった。The crude protein content of the obtained bacterial cells was approximately 77%.
実施例 ８
実施例１と同様な培地、培養条件で、チオバチルス エ
スピー Ａ２ ＡＴＣＣ２５３６４を培養した。Example 8 Thiobacillus sp. A2 ATCC25364 was cultured using the same medium and culture conditions as in Example 1.
３日間培養した後、培養液中のメチルアミン濃度は、０
．００１ｗｔ％以下となった。After culturing for 3 days, the methylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して、菌体を分離回収し、この菌
体を１０５℃で２４時間乾燥して、培養液１１あたり１
．３ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were then dried at 105°C for 24 hours.
．． 3 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約７９％であった。The crude protein content of the obtained bacterial cells was about 79%.
実施例 ９
メチルアミン塩酸塩のかわりに、トリメチルアミン塩酸
塩を用いた他は実施例２と同様な培地、培養条件で、メ
チロバクテリウム オルガノヒラム ＸＸ ＡＴＣＣ
２７８８６を培養した。Example 9 Methylobacterium organohylum XX ATCC was grown using the same medium and culture conditions as in Example 2, except that trimethylamine hydrochloride was used instead of methylamine hydrochloride.
27886 was cultured.
３日間培養した後培養液中のトリメチルアミン濃度は０
．００１ｗｔ％ 以下となった。After culturing for 3 days, the trimethylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５℃で２４時間乾燥して、培養液１１あたり３．
２ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105° C. for 24 hours.
2 g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約８０％であった。The crude protein content of the obtained bacterial cells was approximately 80%.
実施例 １０
メチルアミン塩酸塩のかわりにジメチルアミン塩酸塩を
用いた他は実施例６と同様な培地、培養条件でミクロチ
クルス メタノリカ 微工研菌寄第４４１８号の培養を
行なった。Example 10 Microticulus methanolica Microticulus methanolica No. 4418 was cultured using the same medium and culture conditions as in Example 6, except that dimethylamine hydrochloride was used instead of methylamine hydrochloride.
３日間培養した後、培養液中のジメチルアミン濃度は０
．００１ｗｔ％以下となった。After culturing for 3 days, the dimethylamine concentration in the culture solution was 0.
．． 001wt% or less.
この培養液を遠心分離して菌体を分離回収し、この菌体
を１０５℃で２４時間乾燥して培養液１１あたり２．５
ｇの乾燥菌体を得た。This culture solution was centrifuged to separate and recover the bacterial cells, which were dried at 105°C for 24 hours to give a
g of dried bacterial cells were obtained.
得られた菌体の粗蛋白含量は約８０％であった。The crude protein content of the obtained bacterial cells was approximately 80%.
Claims (1)
ロチクルス属、バラコツカス属またはチオバチルス属に
属し、メチルアミン類を資化しうる細菌を、メチルアミ
ン類を主たる炭素源とする培地に培養し、該培養液から
細菌の菌体を分離することを特徴とする細菌菌体の製造
方法。1. Culture bacteria that belong to the genus Achromobacter, Methylobacterium, Microticulus, Baracoccus, or Thiobacillus and can assimilate methylamines in a medium containing methylamines as the main carbon source, and prepare the culture solution. A method for producing bacterial cells, which comprises separating bacterial cells from.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1398181A JPS5811193B2 (en) | 1981-02-02 | 1981-02-02 | Method for producing bacterial cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1398181A JPS5811193B2 (en) | 1981-02-02 | 1981-02-02 | Method for producing bacterial cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS57129686A JPS57129686A (en) | 1982-08-11 |
| JPS5811193B2 true JPS5811193B2 (en) | 1983-03-01 |
Family
ID=11848395
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1398181A Expired JPS5811193B2 (en) | 1981-02-02 | 1981-02-02 | Method for producing bacterial cells |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5811193B2 (en) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3006453U (en) * | 1994-07-07 | 1995-01-24 | 株式会社栗本鐵工所 | Fastening stop for polyethylene sleeves covering underground pipes |
| US9745489B2 (en) | 2005-07-26 | 2017-08-29 | Knauf Insulation, Inc. | Binders and materials made therewith |
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| US10759695B2 (en) | 2007-01-25 | 2020-09-01 | Knauf Insulation, Inc. | Binders and materials made therewith |
| US10767050B2 (en) | 2011-05-07 | 2020-09-08 | Knauf Insulation, Inc. | Liquid high solids binder composition |
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| US10913760B2 (en) | 2010-05-07 | 2021-02-09 | Knauf Insulation, Inc. | Carbohydrate binders and materials made therewith |
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| US11248108B2 (en) | 2017-01-31 | 2022-02-15 | Knauf Insulation Sprl | Binder compositions and uses thereof |
| US11332577B2 (en) | 2014-05-20 | 2022-05-17 | Knauf Insulation Sprl | Binders |
| US11401204B2 (en) | 2014-02-07 | 2022-08-02 | Knauf Insulation, Inc. | Uncured articles with improved shelf-life |
| US11453780B2 (en) | 2007-01-25 | 2022-09-27 | Knauf Insulation, Inc. | Composite wood board |
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-
1981
- 1981-02-02 JP JP1398181A patent/JPS5811193B2/en not_active Expired
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3006453U (en) * | 1994-07-07 | 1995-01-24 | 株式会社栗本鐵工所 | Fastening stop for polyethylene sleeves covering underground pipes |
| US9745489B2 (en) | 2005-07-26 | 2017-08-29 | Knauf Insulation, Inc. | Binders and materials made therewith |
| US10759695B2 (en) | 2007-01-25 | 2020-09-01 | Knauf Insulation, Inc. | Binders and materials made therewith |
| US11459754B2 (en) | 2007-01-25 | 2022-10-04 | Knauf Insulation, Inc. | Mineral fibre board |
| US10968629B2 (en) | 2007-01-25 | 2021-04-06 | Knauf Insulation, Inc. | Mineral fibre board |
| US11453780B2 (en) | 2007-01-25 | 2022-09-27 | Knauf Insulation, Inc. | Composite wood board |
| US11401209B2 (en) | 2007-01-25 | 2022-08-02 | Knauf Insulation, Inc. | Binders and materials made therewith |
| US10738160B2 (en) | 2010-05-07 | 2020-08-11 | Knauf Insulation Sprl | Carbohydrate polyamine binders and materials made therewith |
| US10913760B2 (en) | 2010-05-07 | 2021-02-09 | Knauf Insulation, Inc. | Carbohydrate binders and materials made therewith |
| US11078332B2 (en) | 2010-05-07 | 2021-08-03 | Knauf Insulation, Inc. | Carbohydrate polyamine binders and materials made therewith |
| US10767050B2 (en) | 2011-05-07 | 2020-09-08 | Knauf Insulation, Inc. | Liquid high solids binder composition |
| US11453807B2 (en) | 2012-04-05 | 2022-09-27 | Knauf Insulation, Inc. | Binders and associated products |
| US11401204B2 (en) | 2014-02-07 | 2022-08-02 | Knauf Insulation, Inc. | Uncured articles with improved shelf-life |
| US11332577B2 (en) | 2014-05-20 | 2022-05-17 | Knauf Insulation Sprl | Binders |
| US10864653B2 (en) | 2015-10-09 | 2020-12-15 | Knauf Insulation Sprl | Wood particle boards |
| US11060276B2 (en) | 2016-06-09 | 2021-07-13 | Knauf Insulation Sprl | Binders |
| US11248108B2 (en) | 2017-01-31 | 2022-02-15 | Knauf Insulation Sprl | Binder compositions and uses thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS57129686A (en) | 1982-08-11 |
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