JPS6258989A - Production of mold of bacterium - Google Patents
Production of mold of bacteriumInfo
- Publication number
- JPS6258989A JPS6258989A JP60198327A JP19832785A JPS6258989A JP S6258989 A JPS6258989 A JP S6258989A JP 60198327 A JP60198327 A JP 60198327A JP 19832785 A JP19832785 A JP 19832785A JP S6258989 A JPS6258989 A JP S6258989A
- Authority
- JP
- Japan
- Prior art keywords
- gas
- hydrogen
- bacterium
- culture
- nitrogen
- 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
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fodder In General (AREA)
Abstract
Description
【発明の詳細な説明】
イ）産業上の利用分野
本発明は、微生物菌体の製造法に関するものであり、よ
り詳しくはキサントバクタ−属に属する酸素耐性水素細
菌を窒素ガスを唯一の窒素源として培養し、これを採取
する微生物菌体の製造法に関するものである。Detailed Description of the Invention A) Industrial Application Field The present invention relates to a method for producing microbial cells, and more specifically, the present invention relates to a method for producing microbial cells. This invention relates to a method for producing microbial cells by culturing and collecting them.
従来、炭酸ガスを炭素源として利用可能μ微生物を培養
して微生物菌体を回収せんとする場合、水素ｆ、［Ｂ菌
が０゛利であると云われている。Conventionally, when attempting to culture μ microorganisms that can use carbon dioxide gas as a carbon source and recover microbial cells, it is said that hydrogen f, [B bacteria have an advantage of 0.
その理由は、原料中に不純物がなくクリーンであるため
、微生物菌体を飼料としてＦｌｌ用する際、η性の問題
が少ないこと、水素細菌の増殖速度が独立栄な細菌の中
で、特に速いことなどが挙げられる。The reason for this is that the raw materials are clean with no impurities, so there are fewer problems with η when using microbial cells as feed, and the growth rate of hydrogen bacteria is particularly fast compared to other independent bacteria. There are many things that can be mentioned.
口）野点を解決するだめの手段
そこで、本発明者らは先にキサントバクター属に属し、
高濃度酸素条件下で生育できる水素細菌の６１１成方法
を提供した（特公昭５９５１９９５号）が更に創成され
た微生物の有効的な利用方法について研究を重ねた結果
、窒素ガスを唯一の窒素源として生育することに着目し
、本発明を完成させたものである。[Example] A means to solve the problem, the present inventors first discovered that Xanthobacter belongs to the genus Xantobacter.
After providing a method for producing 611 hydrogen bacteria that can grow under high-oxygen conditions (Special Publication No. 5951995), as a result of further research into the effective use of the created microorganisms, we found that nitrogen gas was used as the only nitrogen source. The present invention was completed by focusing on the growth of plants.
ずなわら、キサントバクタ−属に属する水素細菌を酸素
ガス、水素ガス及び炭酸ガスからなる混合ガスの存在下
で培養するにあたり、唯一の窒素源として窒素ガスを用
いて培養することを特徴とする微生物菌体の製造法に関
するものである。A microorganism characterized in that hydrogen bacteria belonging to the genus Xanthobacter are cultured in the presence of a mixed gas consisting of oxygen gas, hydrogen gas and carbon dioxide gas, using nitrogen gas as the sole nitrogen source. This relates to a method for producing bacterial cells.
従来、窒素ガスを唯一の窒素源として、キサントバクタ
−属に属する水素細菌を菌体生産させた報告例はな（本
発明が最初である。Hitherto, there have been no reports of cell production of hydrogen bacteria belonging to the genus Xanthobacter using nitrogen gas as the only nitrogen source (the present invention is the first).
この結果、本発明ではこれまでの無機窒素源に代わるも
のとして空気中の窒素ガスを唯一の窒素源として、有効
に利用できるだけでなく、空気中の酸素ガス、炭酸ガス
をもエネルギー源、炭素源として利用可能になり、微生
物菌体の製造コストを大ｉＪにダウンさせることができ
るものである。As a result, the present invention not only makes it possible to effectively utilize nitrogen gas in the air as the only nitrogen source in place of conventional inorganic nitrogen sources, but also uses oxygen gas and carbon dioxide in the air as an energy source and carbon source. This makes it possible to reduce the manufacturing cost of microbial cells to a large iJ.
木閑の具体的な培養は密閉可能な容器に通常の微生物の
培養に用いられる無機化合物、例えばリン酸塩、マグネ
シウム塩、鉄塩などを添加した水溶液を殺菌後、微生物
を接種する。次いで水素ガス、酸素ガス、炭酸ガス及び
窒素ガスからなる混合ガスを通気し攪拌する。また、こ
れらのガスの組成比は通常、水素ガスｌＯ〜６３％、酸
素ガス１〜２３％、炭酸ガス３〜１０％、窒素ガス１１
〜８３％の範囲で通気される。培養温度は通常２５〜３
７℃、ｐｌ＋は、６．０〜９．５の範囲で培養される。To specifically cultivate Mokkan, microorganisms are inoculated after sterilizing an aqueous solution containing inorganic compounds commonly used for culturing microorganisms, such as phosphates, magnesium salts, and iron salts, in a sealable container. Next, a mixed gas consisting of hydrogen gas, oxygen gas, carbon dioxide gas, and nitrogen gas is aerated and stirred. In addition, the composition ratio of these gases is usually hydrogen gas 10~63%, oxygen gas 1~23%, carbon dioxide gas 3~10%, and nitrogen gas 11%.
Ventilated in the range of ~83%. Culture temperature is usually 25-3
Cultured at 7°C, pl+ ranges from 6.0 to 9.5.
本発明において、好適に用いられる代表的な微生物とし
ては、本発明者らが先に創成したキサントバクタ−Ｙ−
３８株（ＦＥＲＭ Ｐ−６２７４）が挙げられる。In the present invention, a representative microorganism suitably used is Xantobacter Y-
38 strains (FERM P-6274) are listed.
本菌株の蘭学的性質は、以下に示すとおりである。The orchidological properties of this strain are as shown below.
ａ）形態
（１）細胞の形及び大きさ、肉汁寒天培地３０°Ｃｌ２
Ｏ間培養
０．６〜０．８ Ｘｌ、５〜３．０ μの稈、菌、直状
または曲状
（２）細胞の多形成、７０間培養で短桿菌となる。a) Morphology (1) Cell shape and size, broth agar medium 30°Cl2
Cultured for 0.6-0.8 Xl, culm of 5-3.0 μ, fungus, polymorphic of straight or curved (2) cells, becomes short rod after cultured for 70 days.
（３）運動性なし
く４）胞子形成なし
く５）ダラム陰性
（６）非抗酸性
（７）コハク酸添加培地で培役すると分岐細胞がみられ
る。(3) No motility 4) No sporulation 5) Durham negative (6) Non-acid-fast (7) Branched cells are observed when cultured in succinic acid-added medium.
（８）貯蔵物質としてポリ−β−ハイドロキシ酪酸（Ｐ
ＨＢ）を蓄積する。(8) Poly-β-hydroxybutyric acid (P
HB).
ｂ）生育状態
（１）肉汁寒天平板培養・黄色、平滑、光沢あり、円形
のコロニーを形成する。色素の拡散なし
く２）肉ｔ）−寒天斜面’３養：肉汁寒天平板培養と同
し
く３）肉汁液体培養・表面発゛Ｕなし、粘質物生成のた
め培養液の粘性かや＼増加する。b) Growth status (1) Broth agar plate culture - Yellow, smooth, shiny, forming circular colonies. No diffusion of pigment 2) Meat t) - Agar slope '3 cultivation: Same as meat juice agar plate culture 3) Meat juice liquid culture / No surface development, viscosity of culture solution increases due to mucilage production .
（４）肉汁ゼラチン穿刺培養：生育せず、液化もなし
く５）リトマル・ミルク：凝固、液化なし、長時間培養
するとアルカリ性となる。(4) Meat juice gelatin puncture culture: No growth, no liquefaction. 5) Lithomal milk: No coagulation, no liquefaction. Becomes alkaline when cultured for a long time.
Ｃ）生理学的性質
（１）硝酸塩の還元
（２）脱窒反応 −
（３）ＭＲテスト −
（４）ＶＰテスト −
（５）インドール生成 −
（６）硫化水素の生成 −
（７）デンプンの加水分解 −
（８）クエン酸の利用
（Ｋ ｏｓｅｒ培地、Ｃｈｒ ｉ ｓ ｔｅｎｓｅｎ培
地）＋（９）無機窒素源の利用
（硝酸塩、アンモニウム塩） ＋（ｌＯ）色素の
生成 非水溶性黄色色素生成（１１）ウレアーゼ
（１２）オキソダーゼ ＋（１３）カタラ
ーゼ 士（１４）生育の範囲
ｐｌ＋ ６．０〜９．５温度２５〜３７℃
（１５）酸素に対する態度 好気性（１６）○−Ｆ
テスト
（ｌｌｕｇｈ Ｌｅｉｆｓｏｎ法による） −（
１７）ｔＱｉＹｉからの酸及びガスの生成 ＝（＋
８）無機化合物のみの固体または液体培地中で、水素ガ
ス、酸素ガス、炭酸ガスの共存下で生育
（１９）メタノール、エタノール、ｎ−プロパツール、
ｎ−ブクノール、ギ酸、酢酸、プロピオン酸、コハク酸
、グルコン酸、などのアルコール類、有機酸類を唯一の
炭素源として生育
（２０）窒素ガス固定能力あり
（２１）水不溶性のカロチノイド色素、ゼアキサンチン
・ディラムノシトを生成する。C) Physiological properties (1) Nitrate reduction (2) Denitrification reaction - (3) MR test - (4) VP test - (5) Indole production - (6) Hydrogen sulfide production - (7) Starch hydration Decomposition - (8) Utilization of citric acid (Koser medium, Chrystensen medium) + (9) Utilization of inorganic nitrogen sources (nitrates, ammonium salts) + (1O) production of pigment Production of water-insoluble yellow pigment (11 ) Urease (12) Oxodase + (13) Catalase (14) Growth range
pl+ 6.0-9.5 Temperature 25-37℃ (15) Attitude towards oxygen Aerobic (16)○-F
Test (by lugh Leifson method) −(
17) Production of acid and gas from tQiYi = (+
8) Growth in a solid or liquid medium containing only inorganic compounds in the coexistence of hydrogen gas, oxygen gas, and carbon dioxide gas (19) Methanol, ethanol, n-propanol,
Grows using alcohols and organic acids such as n-bucnol, formic acid, acetic acid, propionic acid, succinic acid, and gluconic acid as the sole carbon source (20) Capable of fixing nitrogen gas (21) Water-insoluble carotenoid pigment, zeaxanthin. Generate dirhamnocyto.
以上の菌学的性質から、バージエイズ マニュアル オ
ブ ンスティマティク バクテリオロン−Ｖｏｌ １、
（１９８４） （Ｂｅｒｇｅｙ’ｓ Ｍａｎｕａｌ
ｏｆＳｙｓｔｅｍａｔｉｃ ［３ａｃｔｅｒｉｏ
ｌｏｇｙ Ｖｏｌ Ｉ （１９８４） ）に徴して
検討した結果、本菌株をキサントバクタ−・オートトロ
フィカスと同定した。Based on the above mycological properties, the Burgess Manual of Stymatic Bacteriolone-Vol. 1,
(1984) (Bergey's Manual
of Systematic [3acterio
As a result of an investigation based on the results of this study, the strain was identified as Xantobacter autotrophicus.
ハ）実施例
実施例１
−ＶサントバクターＹ−３８株（ＦＥＲＭ Ｐ−［１
７２４）をフラスコ中にて培養した。c) Examples Example 1 -V Santobacter Y-38 strain (FERM P-[1
724) was cultured in flasks.
培地組成は茎溜水１βにリン酸−カリウム３００■、リ
ン酸二カリウム４００ ｏｖ、硫酸マグネシウム２００
■、硫酸第１鉄５０ｍｇ、硫酸亜鉛０．０５■、モリブ
テン酸ナトリウム０．１■、硫酸ニッケル０．２ Ｎを
含むものでｐＨは７．０．１７！フラスコ中の培地Ｉｏ
ｎ １２ニーｌ−４Ｊ−７１バクターＹ−３８株を乾燥
重量として０．３■相相当接接し、フラスコ内の気相を
Ｈ２２０％＋０２４％十Ｃ０２１０％十Ｎ２６６％のｌ
昆合ガスで置換し、３５℃で振盪培養した。The medium composition is 1β of stem water, 300 μl of potassium phosphate, 400 ov of dipotassium phosphate, and 200 μl of magnesium sulfate.
■ Contains 50 mg of ferrous sulfate, 0.05 ■ of zinc sulfate, 0.1 ■ of sodium molybutate, and 0.2 N of nickel sulfate, and has a pH of 7.0.17! Medium Io in flask
n 12 Neal L-4J-71 Bacter Y-38 strain was brought into contact with a dry weight of 0.3 μ phase, and the gas phase in the flask was mixed with H20% + 024% + C02 10% + N266% L
The mixture was replaced with konpo gas and cultured with shaking at 35°C.
１０毎にフラスコ内の気相を同し組成の混合ガスで置換
した。８９時間培養後の菌体濃度は乾燥型！ｆｉ４．５
ｇ／βとなった。この菌体生産成績はアルカリゲネス・
オー１〜二Ｉ・リフィカンスＡ Ｊ ３９４２のジャー
焙１例（特公昭５８−９６７２）を約３倍上回るもので
ある。Every 10 minutes, the gas phase in the flask was replaced with a mixed gas of the same composition. The bacterial cell concentration after 89 hours of culture is dry! fi4.5
g/β. This bacterial production result indicates that Alcaligenes
This is approximately three times higher than the jar roasted example of O 1-2 I. Reficans A J 3942 (Special Publication No. 58-9672).
実施例２
キサントバクタ−Ｙ−３８株（ＦＥＲＭ Ｐ−６７２
４）を空気、水素ガス、炭酸ガスの混合ガスを通気する
小型ジャーで培養した。Example 2 Xanthobacter-Y-38 strain (FERM P-672
4) was cultured in a small jar aerated with a mixed gas of air, hydrogen gas, and carbon dioxide gas.
２ｅの小型ガラス製ジャーに実施例１と同し組成の培地
９００ｍ７！を入れ殺菌後、Ｙ−３８株のフラスコでの
前培養液１００ｍ１を接種した。温度３５°Ｃ１ｐｌ＋
７．０、通気’１１１ ／ｍｉｎ 、攪拌速度４００ｒ
ｐｍで培養した。900 m7 of medium with the same composition as Example 1 in a 2e small glass jar! After sterilization, 100 ml of a flask preculture of Y-38 strain was inoculated. Temperature 35°C 1pl+
7.0, ventilation '111/min, stirring speed 400r
Cultured at pm.
通気ガス組成は培養開始時はＨ２１０％＋０２１％＋Ｃ
○ｚ ＋Ｏ％十Ｎ２７９％であり、菌の生育に伴ってＩ
−（２，Ｏ，濃度を徐々に増大させ、最終的には空気７
５％＋８２２０％＋ＣＯ□ ５％とした。Venting gas composition is H210%+021%+C at the start of culture.
○z +O%10N279%, and as the bacteria grow, I
-(2, O, gradually increasing the concentration until finally air 7
5%+8220%+CO□ 5%.
１０５時間培養後の菌体濃度は乾燥重量で７．５ｇ／ρ
となった。この菌体生産成績はアルカリゲネス・オート
ニトリフィカンスＡＪ３９４２のジャー培養例（特公昭
５８−９６７２）を約４倍上回るものである。The bacterial cell concentration after 105 hours of culture was 7.5 g/ρ in terms of dry weight.
It became. This bacterial cell production result is approximately four times higher than the jar culture example of Alcaligenes autonitrificans AJ3942 (Japanese Patent Publication No. 58-9672).
実施例３
キサントバクタ−Ｙ−３８株（ＦＥｒｌＭ Ｐ−６７
２４）を溶存酸素／二度を０．３５ｐｐｍに維持してジ
ャー培面した。実施例２と同し条件（但し攪拌速度６０
０ｒｐｍ）でジャー培養を行った。Example 3 Xanthobacter Y-38 strain (FErlM P-67
24) was cultured in a jar with dissolved oxygen maintained at 0.35 ppm. Same conditions as Example 2 (but stirring speed 60
Jar culture was performed at 0 rpm).
溶存酸素濃度調節計により溶存酸素？二度を０．３５ｐ
ｐｍに５：Ｉｋ持した。通気ガス組成は培養開始時はＨ
，１３％→−Ｏｚ １ ％ ＋ Ｃ０２３％十Ｎ２８
３％、菌の生育に伴ってＨ２１ζ度を徐々に増大させ培
養終了時はト１２６３％１０□２３％→−Ｃｏ２３％−
←Ｎｚ１１％であった。最終菌体濃度は７２時間１＆養
後に乾燥重量として１３．５ｇ ／ ｊ！となった。こ
の菌体生産成績はアルカリゲネス・オートニトリフィカ
ンスＡＪ３９４２のジャー培養例（特公昭５８−９７６
２）を約６倍上回るものである。Dissolved oxygen by dissolved oxygen concentration controller? 0.35p for twice
I had 5:Ik at pm. The aeration gas composition is H at the start of culture.
,13%→-Oz 1% + C023%10N28
3%, and the H21ζ degree gradually increases as the bacteria grow, and at the end of the culture, it becomes 1263% 10□23% → -Co23%-
←Nz was 11%. The final bacterial cell concentration was 13.5 g/j as dry weight after 72 hours of incubation. It became. This bacterial cell production result is an example of jar culture of Alcaligenes autonitrificans AJ3942 (Special Publication No. 58-976).
2) is approximately six times higher.
官庁出願
手υε令甫正書（方式）
％式％
１、事件の表示 昭和６０年特許願第 １９８３２
７号４、指定代理人
６、補正により増加する発明の数 な し７、補正の
対象
明細書全文Official application form υεReifu official document (method) % formula % 1. Indication of case 1985 Patent Application No. 19832
No. 7 No. 4, Designated Agent 6, Number of inventions increased by amendment None 7, Full text of the specification subject to amendment
Claims (1)
ス及び炭酸ガスから成る混合ガスの存在下で培養するに
あたり、唯一の窒素源として窒素ガスを用いて培養し、
これを採取することを特徴とする微生物菌体の製造法。In culturing hydrogen bacteria belonging to Xanthobacter in the presence of a mixed gas consisting of oxygen gas, hydrogen gas and carbon dioxide gas, culturing using nitrogen gas as the sole nitrogen source,
A method for producing microbial cells, which comprises collecting the cells.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60198327A JPS6258989A (en) | 1985-09-07 | 1985-09-07 | Production of mold of bacterium |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP60198327A JPS6258989A (en) | 1985-09-07 | 1985-09-07 | Production of mold of bacterium |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6258989A true JPS6258989A (en) | 1987-03-14 |
| JPH0378991B2 JPH0378991B2 (en) | 1991-12-17 |
Family
ID=16389261
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP60198327A Granted JPS6258989A (en) | 1985-09-07 | 1985-09-07 | Production of mold of bacterium |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6258989A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114599779A (en) * | 2019-10-29 | 2022-06-07 | 太阳食物有限公司 | Strains and methods for single-cell protein or biomass production |
| JP2025022882A (en) * | 2010-04-27 | 2025-02-14 | キベルディ インコーポレイテッド | Use of oxyhydrogen microorganisms for the recovery and conversion of non-photosynthetic carbon from inorganic and/or C1 carbon sources into useful organic compounds |
-
1985
- 1985-09-07 JP JP60198327A patent/JPS6258989A/en active Granted
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2025022882A (en) * | 2010-04-27 | 2025-02-14 | キベルディ インコーポレイテッド | Use of oxyhydrogen microorganisms for the recovery and conversion of non-photosynthetic carbon from inorganic and/or C1 carbon sources into useful organic compounds |
| CN114599779A (en) * | 2019-10-29 | 2022-06-07 | 太阳食物有限公司 | Strains and methods for single-cell protein or biomass production |
| JP2023500430A (en) * | 2019-10-29 | 2023-01-06 | ソーラー フーズ オサケユイチア | Strains and methods for single cell protein or biomass production |
| US20240093141A1 (en) * | 2019-10-29 | 2024-03-21 | Solar Foods Oy | Strains and processes for single cell protein or biomass production |
| JP2024056855A (en) * | 2019-10-29 | 2024-04-23 | ソーラー フーズ オサケユイチア | Bacterial strains and methods for single cell protein or biomass production |
| CN114599779B (en) * | 2019-10-29 | 2024-05-24 | 太阳食物有限公司 | Strains and methods for single cell protein or biomass production |
| US12600940B2 (en) * | 2019-10-29 | 2026-04-14 | Solar Foods Oyj | Strains and processes for single cell protein or biomass production |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0378991B2 (en) | 1991-12-17 |
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