JPS62236484A - Immobilization of microorganism - Google Patents
Immobilization of microorganismInfo
- Publication number
- JPS62236484A JPS62236484A JP61076631A JP7663186A JPS62236484A JP S62236484 A JPS62236484 A JP S62236484A JP 61076631 A JP61076631 A JP 61076631A JP 7663186 A JP7663186 A JP 7663186A JP S62236484 A JPS62236484 A JP S62236484A
- Authority
- JP
- Japan
- Prior art keywords
- extract
- nutrient
- carrier
- acetobacterium
- growth
- 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
- 244000005700 microbiome Species 0.000 title claims abstract description 19
- 241001468161 Acetobacterium Species 0.000 claims abstract description 14
- 235000015097 nutrients Nutrition 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- 230000003100 immobilizing effect Effects 0.000 claims abstract description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 15
- 239000000284 extract Substances 0.000 abstract description 11
- 239000001888 Peptone Substances 0.000 abstract description 7
- 108010080698 Peptones Proteins 0.000 abstract description 7
- 229940041514 candida albicans extract Drugs 0.000 abstract description 7
- 235000019319 peptone Nutrition 0.000 abstract description 7
- 239000012138 yeast extract Substances 0.000 abstract description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 abstract description 6
- 244000061456 Solanum tuberosum Species 0.000 abstract description 4
- 235000002595 Solanum tuberosum Nutrition 0.000 abstract description 4
- -1 malt extract Substances 0.000 abstract description 4
- 235000013372 meat Nutrition 0.000 abstract description 4
- 239000000203 mixture Substances 0.000 abstract description 4
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 abstract description 3
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 abstract description 3
- 239000001569 carbon dioxide Substances 0.000 abstract description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 abstract description 3
- 239000001257 hydrogen Substances 0.000 abstract description 3
- 229910052739 hydrogen Inorganic materials 0.000 abstract description 3
- 235000010413 sodium alginate Nutrition 0.000 abstract description 3
- 239000000661 sodium alginate Substances 0.000 abstract description 3
- 229940005550 sodium alginate Drugs 0.000 abstract description 3
- 229920001817 Agar Polymers 0.000 abstract description 2
- ZNOZWUKQPJXOIG-XSBHQQIPSA-L [(2r,3s,4r,5r,6s)-6-[[(1r,3s,4r,5r,8s)-3,4-dihydroxy-2,6-dioxabicyclo[3.2.1]octan-8-yl]oxy]-4-[[(1r,3r,4r,5r,8s)-8-[(2s,3r,4r,5r,6r)-3,4-dihydroxy-6-(hydroxymethyl)-5-sulfonatooxyoxan-2-yl]oxy-4-hydroxy-2,6-dioxabicyclo[3.2.1]octan-3-yl]oxy]-5-hydroxy-2-( Chemical compound O[C@@H]1[C@@H](O)[C@@H](OS([O-])(=O)=O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H]2OC[C@H]1O[C@H](O[C@H]1[C@H]([C@@H](CO)O[C@@H](O[C@@H]3[C@@H]4OC[C@H]3O[C@H](O)[C@@H]4O)[C@@H]1O)OS([O-])(=O)=O)[C@@H]2O ZNOZWUKQPJXOIG-XSBHQQIPSA-L 0.000 abstract description 2
- 239000008272 agar Substances 0.000 abstract description 2
- 229920002401 polyacrylamide Polymers 0.000 abstract description 2
- 230000000813 microbial effect Effects 0.000 abstract 2
- 239000006285 cell suspension Substances 0.000 abstract 1
- 239000003795 chemical substances by application Substances 0.000 abstract 1
- 238000001879 gelation Methods 0.000 abstract 1
- 230000035755 proliferation Effects 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 description 24
- 239000000243 solution Substances 0.000 description 9
- 239000000463 material Substances 0.000 description 8
- 241000093709 Acetobacterium sp. Species 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 235000015193 tomato juice Nutrition 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 150000003722 vitamin derivatives Chemical class 0.000 description 3
- ALYNCZNDIQEVRV-UHFFFAOYSA-N 4-aminobenzoic acid Chemical compound NC1=CC=C(C(O)=O)C=C1 ALYNCZNDIQEVRV-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- 235000010410 calcium alginate Nutrition 0.000 description 2
- 239000000648 calcium alginate Substances 0.000 description 2
- 229960002681 calcium alginate Drugs 0.000 description 2
- OKHHGHGGPDJQHR-YMOPUZKJSA-L calcium;(2s,3s,4s,5s,6r)-6-[(2r,3s,4r,5s,6r)-2-carboxy-6-[(2r,3s,4r,5s,6r)-2-carboxylato-4,5,6-trihydroxyoxan-3-yl]oxy-4,5-dihydroxyoxan-3-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylate Chemical compound [Ca+2].O[C@@H]1[C@H](O)[C@H](O)O[C@@H](C([O-])=O)[C@H]1O[C@H]1[C@@H](O)[C@@H](O)[C@H](O[C@H]2[C@H]([C@@H](O)[C@H](O)[C@H](O2)C([O-])=O)O)[C@H](C(O)=O)O1 OKHHGHGGPDJQHR-YMOPUZKJSA-L 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 229910052500 inorganic mineral Inorganic materials 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000011707 mineral Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 description 1
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 1
- AGBQKNBQESQNJD-SSDOTTSWSA-N (R)-lipoic acid Chemical compound OC(=O)CCCC[C@@H]1CCSS1 AGBQKNBQESQNJD-SSDOTTSWSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 241000511222 Acetobacterium wieringae Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- JOYRKODLDBILNP-UHFFFAOYSA-N Ethyl urethane Chemical compound CCOC(N)=O JOYRKODLDBILNP-UHFFFAOYSA-N 0.000 description 1
- LKDRXBCSQODPBY-AMVSKUEXSA-N L-(-)-Sorbose Chemical compound OCC1(O)OC[C@H](O)[C@@H](O)[C@@H]1O LKDRXBCSQODPBY-AMVSKUEXSA-N 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- 239000004372 Polyvinyl alcohol Substances 0.000 description 1
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- AGBQKNBQESQNJD-UHFFFAOYSA-N alpha-Lipoic acid Natural products OC(=O)CCCCC1CCSS1 AGBQKNBQESQNJD-UHFFFAOYSA-N 0.000 description 1
- 229960004050 aminobenzoic acid Drugs 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229960001305 cysteine hydrochloride Drugs 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 239000003349 gelling agent Substances 0.000 description 1
- 235000019136 lipoic acid Nutrition 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- MGFYIUFZLHCRTH-UHFFFAOYSA-N nitrilotriacetic acid Chemical compound OC(=O)CN(CC(O)=O)CC(O)=O MGFYIUFZLHCRTH-UHFFFAOYSA-N 0.000 description 1
- 229940014662 pantothenate Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 229920002451 polyvinyl alcohol Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- ZUFQODAHGAHPFQ-UHFFFAOYSA-N pyridoxine hydrochloride Chemical compound Cl.CC1=NC=C(CO)C(CO)=C1O ZUFQODAHGAHPFQ-UHFFFAOYSA-N 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000002344 surface layer Substances 0.000 description 1
- 229960002663 thioctic acid Drugs 0.000 description 1
- 230000002747 voluntary effect Effects 0.000 description 1
Landscapes
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Distillation Of Fermentation Liquor, Processing Of Alcohols, Vinegar And Beer (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、二酸化炭素と水素から酢酸を生産するアセト
バクテリウム属の微生物の固定化方法に関する
（従来技術）
現在、固定化微生物を利用する連続発酵システムは、工
業プロセスにおいて大きな注［１を浴びている。このシ
ステムは、従来より行なわれている回分発酵に比べてエ
ネルギー的コストが高効率であるプロセスとして発展し
ている。イの中でも特に、菌体分離等の工程でエネルギ
ーコストが低減されるメリットがある。Detailed Description of the Invention (Industrial Application Field) The present invention relates to a method for immobilizing Acetobacterium microorganisms that produce acetic acid from carbon dioxide and hydrogen (prior art) Currently, immobilized microorganisms are used. Continuous fermentation systems have received great attention [1] in industrial processes. This system has been developed as a process with high energy cost efficiency compared to conventional batch fermentation. In particular, it has the advantage of reducing energy costs in processes such as bacterial cell isolation.
（発明が解決しようとしている問題点）しかし、 ＩＬ
ｊ定化激化微生物いても、固定化物の表層では基質との
接触効率が高く、微生物の増殖が著しい反面、拡散抵抗
のため基質は固定化物内部に入りにクク、微生物の増殖
が抑制され、固定化物外へ流出する国体量が非常に高く
なってくる。(The problem that the invention is trying to solve) However, IL
Even if there are microorganisms, the surface layer of the immobilized material has a high contact efficiency with the substrate, and the microorganisms proliferate rapidly. However, due to diffusion resistance, the substrate cannot enter the interior of the immobilized material, suppressing the growth of microorganisms and causing the immobilization. The amount of national polity flowing out of the world is becoming extremely high.
さらには、微生物の大きさと固定化マトリックスでの孔
径との関係から、バクアリアの場合特に流出が顕著であ
り、固定化物よりの微生物の流出防止が強く望まれてい
る。また、アセトバクテリウム属に屈する微生物による
酢酸の連続発酵システムのための固定化方法は、全く検
討されていなかった。Furthermore, due to the relationship between the size of the microorganism and the pore diameter of the immobilized matrix, the outflow is particularly pronounced in the case of bacteria, and there is a strong desire to prevent the outflow of microorganisms from the immobilized material. In addition, an immobilization method for a continuous fermentation system of acetic acid using microorganisms belonging to the genus Acetobacterium has not been studied at all.
（問題点を解決するための手段）
本発明者らは、アセトバクテリウム属に属する微生物を
用いて酢酸を連続的に製造する方法の研究において、固
定化物より流出する菌体Ｍを防止することを検討した結
束、固定化物内に酵母エキスなどの生育促進栄養物を添
加すると流出を防止できることを見い出し１本発明を完
成した。すなわら１本発明はアセトバクテリウム属の生
育を促進さＵる栄養物である。酵母エキス、ジャガイモ
エキス、麦芽エキス、ペプトン、肉エキス、トマトジュ
ースエキス、ミルクペプトンなどを固定化物の中に添加
し、その内部での増殖を高め、更に流出することを押え
１分離処理への負荷をかけないアセトバクテリウム属の
微生物の固定化方法である。(Means for Solving the Problems) In researching a method for continuously producing acetic acid using microorganisms belonging to the genus Acetobacterium, the present inventors aimed to prevent bacterial cells M from flowing out from an immobilized product. The present invention was completed based on the discovery that adding growth-promoting nutrients such as yeast extract to the bound and immobilized material can prevent leakage. Specifically, the present invention is a nutrient that promotes the growth of Acetobacterium. Yeast extract, potato extract, malt extract, peptone, meat extract, tomato juice extract, milk peptone, etc. are added to the immobilized material to increase its internal growth and prevent it from flowing out. 1. Load on the separation process This is a method for immobilizing Acetobacterium microorganisms without applying heat.
本発明で使用する微生物としては、アセトバクテリウム
属に属する二酸化炭素と水素を資化することのできる微
生物であればいずれでも良いが。The microorganism used in the present invention may be any microorganism belonging to the genus Acetobacterium that can assimilate carbon dioxide and hydrogen.
例えばアセトバクテリウム・ウツディ（Ａｃｅｔｏｂａ
ｃｔｅｒｉｕｍ ｗｏｏｄｉｉ）、アセトバクテリウ
ム・ライリンガ−（Ａｃｅｔｏｂａｃｔｅｒｉｕｍ
ｗｉｅｒｉｎｇａｅ）、アセトバクテリウム・カルビノ
リカム（Ａ ＣＯｊ Ｏｂ ａ Ｃｔｅｒｉｕｍ ｃ
ａｒｂｉｎｏｌｉｃｕｍ）、アセトバクテリウム・エス
ピーＮｏ、４４６ （Ａｃｅｔｏｂａｃｔｅｒｉｕｍ
ＳＰ、）ＦＥＲＭＰ−７０１７などを挙げることがで
きる。For example, Acetobacterium uthudi (Acetobacterium utsudi)
cterium woodii), Acetobacterium reilinger (Acetobacterium
wieringae), Acetobacterium carbinoricum (A COj Ob a Cterium c
arbinolicum), Acetobacterium sp. No. 446 (Acetobacterium sp.
SP, ) FERMP-7017 and the like.
本発明に於いて用いる菌体は９国体を嫌気条件下で培養
した摂、培養液から分離した菌体そのまま使用すること
が好ましい。In the present invention, it is preferable that the bacterial cells used in the present invention be those isolated from the culture solution obtained by culturing the nine-keptidium nigra under anaerobic conditions and used as they are.
本発明で用いる固定化１１休どしては、主どして包括方
法に用いられる担体であればいずれでも良いが、例えば
アルギン酸少１−リウム、カッパーカラギーナン、ポリ
アクリルアミド、光硬化樹脂。For immobilization 11 used in the present invention, any carrier that is mainly used in the entrapment method may be used, such as oligolium alginate, kappa carrageenan, polyacrylamide, and photocurable resin.
ウレタンプレポリマー、寒天、ポリビニルアルコール、
ペクチンなどが挙げられる。Urethane prepolymer, agar, polyvinyl alcohol,
Examples include pectin.
本発明で用いる生育促進栄養物としては、アセトバクゾ
リウム属の微生物の生育を促進させる栄養物であればい
ずれでも良いが、具体的には酵母エキス、ジャガイモエ
キス、麦芽エキス、ペプトン、肉エキス、トマトジュー
スエキス、ミルクペプトンなどである。The growth-promoting nutrients used in the present invention may be any nutrients that promote the growth of microorganisms of the genus Acetobaczolium, but specific examples include yeast extract, potato extract, malt extract, peptone, and meat extract. , tomato juice extract, milk peptone, etc.
固定化方法としては、菌体または菌体Ｉｔ！！濁液に生
育促進栄養物を添加し、そこに前述の担体を加えてゲル
化剤により包括固定化を行なう、菌体に対する生育促進
栄養物質の添加量は、菌体乾燥重量グ当り０．０５ｆｆ
以上が適している。好ましくは０．０５〜０．２ｆｆで
ある。 担体１１１］ＩトＬ、でハ物質により異なるが
２通常は１．５〜３．０％のものである０通常アルギン
酸ナトリウムの場合は１゜用すると有用である。As an immobilization method, bacterial cells or bacterial cells It! ! A growth promoting nutrient is added to the suspension, the above-mentioned carrier is added thereto, and entrapping immobilization is carried out using a gelling agent.The amount of the growth promoting nutrient added to the bacterial cells is 0.05 ff per dry weight of the bacterial cells.
The above is suitable. Preferably it is 0.05 to 0.2ff. Carrier 111] I and L, depending on the substance, is usually 1.5 to 3.0%. Usually, it is useful to use 1° in the case of sodium alginate.
（発明の効果）
本発明の特徴は、固を化物の中にアセトバクテリウム属
の生育を促進さぼる栄養物である酵母エキス、ジャガイ
モエキス、麦芽エキス、ペプトン。(Effects of the Invention) The present invention is characterized by containing yeast extract, potato extract, malt extract, and peptone, which are nutrients that promote and inhibit the growth of Acetobacterium in solid compounds.
肉エキス、トマトジュースエキス、ミルクペプトンを添
加することにより、その固定化物の中で十分増殖させ、
内部から流出する菌体量を少なく押えることができる。By adding meat extract, tomato juice extract, and milk peptone, it can be sufficiently grown in the immobilized material.
It is possible to suppress the amount of bacterial cells flowing out from the inside.
このことにより１分離操作（菌体除去）の時間。This reduces the time required for one isolation operation (removal of bacterial cells).
並びに回転動力を低（保ちながら、連続的な操作が可能
となり、エネルギーコストを大幅に低減することが可能
となる。以下１本発明を実施例より説明する。In addition, continuous operation is possible while keeping the rotational power low, and energy costs can be significantly reduced.The present invention will be explained below with reference to examples.
（実施例）
炭素源として、１％ソルボースを添加した第１表の培地
でアセトバクテリウム・エスピーＮｏ。(Example) Acetobacterium sp. No. was grown in the medium shown in Table 1 to which 1% sorbose was added as a carbon source.
４４６を嫌気的に培養し集菌した後、０．１％酵母エキ
ス水溶液に懸濁する。その懸濁液１容に対して１容の３
％アルギン酸ナトリウム水溶液を加え、十分混合する。After culturing 446 anaerobically and collecting the bacteria, they are suspended in a 0.1% yeast extract aqueous solution. 1 volume of 3 to 1 volume of the suspension
% sodium alginate aqueous solution and mix thoroughly.
その混合液を２２％塩化カルシウム水溶液に滴下し１粒
径２ａｍのアルギン酸カルシウム包括固定化アヒトバク
デリウムエスビーＮｏ、４４６を得た。この調製した固
定化菌体４ｄに対して１６ｄの培地を添加し、３０℃、
１８０回転振盪により培養を行なった。培地組成は。The mixed solution was dropped into a 22% calcium chloride aqueous solution to obtain Ahitobacterium S.B. No. 446 entrapping and immobilizing calcium alginate with a particle size of 2 am. 16 d of medium was added to 4 d of the prepared immobilized bacterial cells, and the mixture was heated at 30°C.
Culture was performed by shaking at 180 rpm. What is the medium composition?
表に示した。固定化物内と固定化物外へ流出した菌体ｍ
の経時変化を図に示した。培養終了時の培養液中の菌体
濃度は、培養液１ 当り０．０２ｇと低く、流出菌体量
は菌体濃度比で０．０７倍であった。Shown in the table. Bacterial cells flowing into and out of the immobilized material
The figure shows the change over time. The bacterial cell concentration in the culture solution at the end of the culture was as low as 0.02 g per culture solution, and the amount of bacterial cells flowing out was 0.07 times the bacterial cell concentration ratio.
（比較例）
アルギン酸カルシウムゲル内に栄養物を添加せず、アセ
トバクゾリウム・エスピーＮｏ、４４６を固定化し、実
施例と同様に培養を行なった場合。(Comparative example) A case where Acetobaczolium sp. No. 446 was immobilized in calcium alginate gel without adding nutrients, and cultured in the same manner as in the example.
培養終了時の培養液中の菌体濃度は培養液１ 当り０．
１３ｇと高く、流出菌体量は菌体濃度比で０．６１倍で
あった。The bacterial cell concentration in the culture solution at the end of the culture is 0.0% per culture solution.
It was high at 13 g, and the amount of bacterial cells flowing out was 0.61 times the bacterial cell concentration ratio.
Ｂｔ口法）
流出菌体量とゲル内の菌体濃度の比を菌体′ｍ度比とす
る。Bt method) The ratio of the amount of bacterial cells flowing out and the concentration of bacterial cells in the gel is defined as the bacterial cell ratio.
ノ８養終了時の液中の菌体１１度
１（ｄｒｙ ｃａｌｌｓ）／
（ｌｉｑｕｉｄ））
菌体′ａ度比−□
培養終了時の固定化物内の菌体量
度（ｇ＜ｄｒｙ ｃｅｌｌｓ）／
（ｌｉｑｕｉｄ＞）
４、図の簡単な、ｉ？２明
実施例に基づいて培養を行なった時の菌体濃度と酢酸生
産の経時変化の図
第１表
０．１％レザズリン １ 成１０％Ｎ
ｉ１４ １０ ｄｌＭ ＫＨ
２ＰＯ４
（Ｐｌ−１７，０＞ ５ ｍｉ！２０％Ｍ
＜ｉｓｏ ７Ｈ２００，５２ｄビタミン溶液
２０ ｄミネラル溶液
４０ ｒｄシスティン塩酸（１水塩）
０．５ ｇＮ ａ ２ Ｓ
０ 、２ ｂ ’ＩＮ ａ ｌ−Ｉ Ｃ０３１
０’、ｉ
酵母エキス ０．２ｆｆ０．０６
％ブロムエタン
スルホン１１１１１１１ｄ
Ｐ＋−４７，８
ビタミン溶液組成 （ｄ／ ）ビデオ
ン ２葉酸
２
ピリドキシン塩酸 １０ヂアミン塩酸
５リボフラビン
５
ニコチン酸 ５
パントテン酸Ｃａ ５ビタミン［３１
２ｏ、ｏｉ
Ｐ−アミノ安息香酸 ５
チオクト酸 １ミネラル溶液
（ｇ／ ）ニトリロ３酢酸
０．２５Ｍ ｎ Ｓ （） ・ ４
ト１２ 。8 Bacterial cells in the liquid at the end of cultivation 11 degrees 1 (dry calls) / (liquid)) Bacterial cell 'a ratio - □ Amount of bacterial cells in the immobilized material at the end of cultivation (g<dry cells) / ( liquid>) 4. Simple i? Table 1: Diagram of changes in bacterial cell concentration and acetic acid production over time when culturing was carried out based on Example 2. 0.1% resazurin 1 10% N
i14 10 dlM KH
2PO4 (Pl-17,0> 5 mi!20%M
<iso 7H200,52d vitamin solution
20 d mineral solution
40 rd cysteine hydrochloride (monohydrate)
0.5 gN a2S
0, 2 b'IN a l-I C031
0', i Yeast extract 0.2ff0.06
% Bromoethanesulfone 1111111d P+-47,8 Vitamin solution composition (d/) Videon 2 folic acid
2 Pyridoxine hydrochloric acid 10diamine hydrochloric acid
5 Riboflavin
5 Nicotinic acid 5 Ca pantothenate 5 Vitamin [31
2o, oi P-aminobenzoic acid 5 Thioctic acid 1 Mineral solution
(g/) Nitrilotriacetic acid
0.25M n S () ・ 4
12.
４ ０、２８
ＮａＣＩ ０．５ＦｅＳＯ−
７Ｈ２０
４０、０５
ＣａＣ１−６８２０
２０、０９
ｃａｃ Ｉ ・２Ｈ２０
２０、０７
ＺｎＳＯ−７１ゴ 、 ０
４ ０、 ０９ＣａＳ
０４ ０．Ｏ’３ＡＩＫ（Ｓ０４）
２・１２Ｈ２０ｏ、ｏｏ９１−１３ＢＯ４０，００５
ＮａＭｏＯ” ２１−１２０ ０．００６特許
出願人 工業技ｌｆＪ院長
峠ｆ’、’Ｉ（ｌｖｚエノ
手続補正書（自発）
昭和６１年？月１２１］4 0, 28 NaCI 0.5FeSO-
7H20 40, 05 CaC1-6820 20, 09 cac I ・2H20 20, 07 ZnSO-71go, 0 4 0, 09CaS
04 0. O'3AIK (S04)
2・12H20o, oo91-13BO40,005 NaMoO" 21-120 0.006 Patent applicant Industrial Technology lfJ Director Touge f','I (lvz Eno procedural amendment (voluntary) 1985? Month 121]
Claims (1)
と共に固定化することを特徴とする微生物の固定化方法A method for immobilizing microorganisms, characterized by immobilizing microorganisms belonging to the genus Acetobacterium together with growth-promoting nutrients.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61076631A JPS62236484A (en) | 1986-04-04 | 1986-04-04 | Immobilization of microorganism |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP61076631A JPS62236484A (en) | 1986-04-04 | 1986-04-04 | Immobilization of microorganism |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS62236484A true JPS62236484A (en) | 1987-10-16 |
| JPH038199B2 JPH038199B2 (en) | 1991-02-05 |
Family
ID=13610718
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP61076631A Granted JPS62236484A (en) | 1986-04-04 | 1986-04-04 | Immobilization of microorganism |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS62236484A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7029884B2 (en) | 1997-05-29 | 2006-04-18 | Japan Science And Technology Corporation | Carrier for microorganism incubation in which micro-elements and inorganic nutrient salts are diffused |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60120987A (en) * | 1983-12-05 | 1985-06-28 | Kikkoman Corp | Production of immobilized microbial cell or enzyme |
-
1986
- 1986-04-04 JP JP61076631A patent/JPS62236484A/en active Granted
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60120987A (en) * | 1983-12-05 | 1985-06-28 | Kikkoman Corp | Production of immobilized microbial cell or enzyme |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7029884B2 (en) | 1997-05-29 | 2006-04-18 | Japan Science And Technology Corporation | Carrier for microorganism incubation in which micro-elements and inorganic nutrient salts are diffused |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH038199B2 (en) | 1991-02-05 |
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