JPH01243997A - Production of microbial cellulose - Google Patents
Production of microbial celluloseInfo
- Publication number
- JPH01243997A JPH01243997A JP7107188A JP7107188A JPH01243997A JP H01243997 A JPH01243997 A JP H01243997A JP 7107188 A JP7107188 A JP 7107188A JP 7107188 A JP7107188 A JP 7107188A JP H01243997 A JPH01243997 A JP H01243997A
- Authority
- JP
- Japan
- Prior art keywords
- culture
- oxygen
- containing gas
- bubble column
- medium
- 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
- 229920001340 Microbial cellulose Polymers 0.000 title claims description 15
- 238000004519 manufacturing process Methods 0.000 title claims description 7
- 239000007789 gas Substances 0.000 claims abstract description 26
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000001301 oxygen Substances 0.000 claims abstract description 17
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 17
- 238000011218 seed culture Methods 0.000 claims abstract description 10
- 238000012258 culturing Methods 0.000 claims abstract description 6
- 229920002678 cellulose Polymers 0.000 claims abstract description 5
- 239000001913 cellulose Substances 0.000 claims abstract description 5
- 230000003068 static effect Effects 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 239000005720 sucrose Substances 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- IMQLKJBTEOYOSI-UHFFFAOYSA-N Phytic acid Natural products OP(O)(=O)OC1C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C(OP(O)(O)=O)C1OP(O)(O)=O IMQLKJBTEOYOSI-UHFFFAOYSA-N 0.000 claims description 4
- 150000001413 amino acids Chemical class 0.000 claims description 4
- IMQLKJBTEOYOSI-GPIVLXJGSA-N Inositol-hexakisphosphate Chemical compound OP(O)(=O)O[C@H]1[C@H](OP(O)(O)=O)[C@@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@H](OP(O)(O)=O)[C@@H]1OP(O)(O)=O IMQLKJBTEOYOSI-GPIVLXJGSA-N 0.000 claims 2
- 229940068041 phytic acid Drugs 0.000 claims 2
- 235000002949 phytic acid Nutrition 0.000 claims 2
- 239000000467 phytic acid Substances 0.000 claims 2
- 244000235858 Acetobacter xylinum Species 0.000 claims 1
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims 1
- 238000011534 incubation Methods 0.000 claims 1
- 239000001963 growth medium Substances 0.000 abstract description 5
- 244000005700 microbiome Species 0.000 abstract 2
- 150000001875 compounds Chemical class 0.000 abstract 1
- 239000002537 cosmetic Substances 0.000 abstract 1
- 239000002609 medium Substances 0.000 description 11
- 239000000499 gel Substances 0.000 description 8
- 238000003756 stirring Methods 0.000 description 7
- 241000894006 Bacteria Species 0.000 description 5
- 238000004140 cleaning Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 238000005273 aeration Methods 0.000 description 3
- 238000012136 culture method Methods 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000013019 agitation Methods 0.000 description 2
- 238000009423 ventilation Methods 0.000 description 2
- RZVAJINKPMORJF-UHFFFAOYSA-N Acetaminophen Chemical compound CC(=O)NC1=CC=C(O)C=C1 RZVAJINKPMORJF-UHFFFAOYSA-N 0.000 description 1
- 244000283763 Acetobacter aceti Species 0.000 description 1
- 235000007847 Acetobacter aceti Nutrition 0.000 description 1
- PTHCMJGKKRQCBF-UHFFFAOYSA-N Cellulose, microcrystalline Chemical compound OC1C(O)C(OC)OC(CO)C1OC1C(O)C(O)C(OC)C(CO)O1 PTHCMJGKKRQCBF-UHFFFAOYSA-N 0.000 description 1
- 241001662103 Cryptocarya corrugata Species 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 235000021185 dessert Nutrition 0.000 description 1
- 229910001882 dioxygen Inorganic materials 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 229920006015 heat resistant resin Polymers 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 229920000515 polycarbonate Polymers 0.000 description 1
- 239000004417 polycarbonate Substances 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
（産業上の利用分野）
本発明は、ＡＣＯｔＯｂａＣｔＯｒ ａｃｅｔｉ ５
ｕｂｓｐ。DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention
ubsp.
ｘｙｌｉｎｕ−を気泡塔培養（菌の増殖）法と棚段静置
培養（微生物セルロースの生成）法とを組合せることに
よる微生物セルロースの大Ｈｔ生産方法に関し、更に詳
しくは、本発明は、△ＣｅｔＯｂａＣｔＯｒａＣｅｔｉ
５ｕｂｓｐ、 ｘｙｌｉｎｕｍをシード培養し、Ｉｎ
１′ｊ１を具備していない気泡塔培養槽中にシードを
接種してＭｉ含右ガスを通気しながら気泡塔培養し、得
られた菌の増殖した培養液を棚段培養器に分注しで静置
培養することから成る微生物セルロースの産生方法に関
する。ΔCetObaCtOraCeti
5ubsp, xylinum was seeded, and In
Seeds were inoculated into a bubble column culture tank not equipped with 1'j1, and cultured in a bubble column while aerating Mi-containing gas, and the obtained culture solution in which the bacteria grew was dispensed into a tray culture vessel. The present invention relates to a method for producing microbial cellulose, which comprises statically culturing the cellulose.
（先行の技術）
Ａｃｅｔｏｂａｃｔｏｒ ａｃｅｔｉ ５ｕｂｓｐ、
ｘｙｌｉｎｕｍは１１１１１１外にセルロース（微生物
セル［］−ス）をシート状又はゲル状に産生する。この
微生物セルロースは弾性、破断強度等の物性面で非常に
優れたもので、デザート食品をはじめ音響板、特殊シー
ト、化蛙品等の巾広い用途が期待されている。(Prior art) Acetobacter aceti 5ubsp,
In addition to 111111, xylinum produces cellulose (microbial cells) in the form of sheets or gels. This microbial cellulose has excellent physical properties such as elasticity and breaking strength, and is expected to be used in a wide range of applications, including dessert foods, acoustic boards, special sheets, and pet frog products.
このような微生物セルロースの生産は静置培養法で行な
われ、培養液の表面に微生物セルロースのシート状膜が
形成する。従来から行なわれている静置培養法は、Ａ
ｃｃｔｏｂａｃｔｏｒ ａｃｅ口５ｕｂｓｐ。Such production of microbial cellulose is carried out by a static culture method, and a sheet-like film of microbial cellulose is formed on the surface of the culture solution. The conventional static culture method is A.
cctobactor ace mouth 5ubsp.
ｘｙｌｉｎｕｍをシード培養し、培養された種菌をコン
テナーのＪＲ地に接種して約２０日静置培ｍする方法で
あり、微生物セルロースの産生に長期を要する欠点があ
る。This method involves seed culturing M. xylinum, inoculating the cultured seed fungus into the JR soil of a container, and culturing it statically for about 20 days, which has the disadvantage that it takes a long time to produce microbial cellulose.
従って、微生物セルロースの天吊生産及び培養期間の大
巾短縮が求められている。Therefore, there is a need to significantly shorten the period of suspended production and cultivation of microbial cellulose.
培養期間の短縮をする為に、まｆ Ａ ｃｅｔｏｂａｃ
ｔｏｒａｃｃｔｉ ５ｕｂｓｐ、 ｘｙｌｉｎｕｍのシ
ード培養ステップと静置培養ステップの間に通気撹拌培
養ステップを導入する方法が考えられた。In order to shorten the culture period, maf A cetobac
A method of introducing an aeration agitation culture step between the seed culture step and the stationary culture step of P. toracti 5ubsp, xylinum was considered.
しかしながら、通気撹拌培養を採用すると、培養期間は
短縮されるが、培養が進むにつれて攪拌機に産生じた微
生物セルロースがからみついたり、 、培養塔内に微
生物セルロースゲルが付着したりして培養器の洗浄に非
常な時間と労力を必要とする問題点がある。However, when aerated agitation culture is adopted, the culture period is shortened, but as the culture progresses, the microbial cellulose produced in the agitator may become entangled, and the microbial cellulose gel may adhere to the inside of the culture tower, causing the incubator to become damaged. There is a problem that cleaning requires a lot of time and effort.
上述のような欠点を解決し、微生物セルロースを大量生
産するための本発明者等の鋭意研究の結果、通気撹拌培
養槽のかわりに気泡塔培養槽を用いて培養すると高収率
で微生物セルロースを得ることが出来、且つ培養槽内の
洗浄が容易且つ簡単に出来るとともに、培改時間を大巾
に短縮し得ることを見出し、この知見に基づいて本発明
を成すにヱった。As a result of intensive research by the present inventors to solve the above-mentioned drawbacks and mass-produce microbial cellulose, microbial cellulose can be produced in high yield when cultured using a bubble column culture tank instead of an aerated stirring culture tank. The present inventors have found that the culture tank can be easily and easily cleaned, and the culture reformation time can be greatly shortened. Based on this knowledge, the present invention has been completed.
（問題点を解決する為の手段）
本発明の微生物セルロースの産生する方法は、Ａｃｅｔ
ｏｂａｃｔｏｒ ａｃｅｔｉ ５ｕｂｓｐ、 ｘｙｌｉ
ｎｕｍをシード培養し、攪拌機を具備していない気泡塔
培養槽中にシードを接種して酸素含有ガスを通気しなが
ら気泡塔培養し、得られた培養液を棚段培養器に分注し
て静置培養することから成る。(Means for Solving the Problems) The method for producing microbial cellulose of the present invention includes Acet
obactor aceti 5ubsp, xyli
num was seeded and cultured in a bubble column culture tank without a stirrer, and cultured in a bubble column while aerating oxygen-containing gas, and the resulting culture solution was dispensed into a tray culture vessel. It consists of static culture.
本発明のシード培養及び気泡塔培養で使用する培地は、
蔗糖又は砂糖；イーストエクストラクト又は総合アミノ
酸とフィチンＩ！：（ＮＨ４）２Ｓｏ ： ＫＨ２ｐ
ｏ４：及びＭｇ８０４を含有し、１１０〜１２０℃で１
０〜３０分間殺菌されたものである。The medium used in the seed culture and bubble column culture of the present invention is
Sucrose or sugar; yeast extract or general amino acids and phytin I! :(NH4)2So :KH2p
o4: Contains Mg804 and 1 at 110-120℃
It is sterilized for 0 to 30 minutes.
シード培養で使用する培地のｌ）Ｈは３．５〜７．５で
、気泡塔培養で使用する培地′のＩ）Ｈは３．５〜７．
５である。The l)H of the medium used in seed culture is 3.5 to 7.5, and the I)H of the medium ' used in bubble column culture is 3.5 to 7.5.
It is 5.
使用される培地の好ましい組成は、蔗糖２〜１０ｇ／旧
又は砂糖２〜１０９ ／ｄｉ ：イーストエクストラク
ト０．１〜１．０ ｓ／ｄｌ又は総合アミノ酸０．１〜
１．０ ｇ／ｄｌ及びフィスチン酸０，０１〜０．０５
ｓｔ／ｄｉ ：（ＮＨ４）２ ＳＯ２ｏ、１〜２．０９
／ｄｌ：ＫＨ２ＰＯ２０，１〜１．０９／ｄｌ及びＭａ
ＳＯ４０，０１〜０．５ ｇ／ｄｌを含有するものであ
る。The preferred composition of the medium used is: sucrose 2-10 g/di or sugar 2-109/di: yeast extract 0.1-1.0 s/dl or total amino acids 0.1-109/di.
1.0 g/dl and fistic acid 0.01-0.05
st/di: (NH4)2SO2o, 1-2.09
/dl: KH2PO20.1-1.09/dl and Ma
It contains SO40.01 to 0.5 g/dl.
シード培養は、殺菌処理された上記培地にＡｃｅｔｏｂ
ａｃｔｏｒ ａｃｅｔｉ ５ｕｂｓｐ、 ｘｙｌｉｎｕ
−菌を接種し、振盪機上で２０〜３５℃、２４〜９６時
間おこなう。For seed culture, Acetob was added to the above sterilized medium.
actor aceti 5ubsp, xylinu
- Inoculate the bacteria and incubate on a shaker at 20-35°C for 24-96 hours.
気泡塔培養槽は、菌の増殖にともなう培養槽内部へのゲ
ルの付着の防止の為に、培養槽内部には撹拌棒や撹拌羽
を用いるような撹拌装置を具備しない代わりに、その底
部に培養時のＭ素供給と内容物の撹拌を目的とした酸素
含有ガスの導入機構を具備している。培養槽上部には導
入ガスの排出し］が、底部には培養液の取出口が設けら
れている。In order to prevent gel from adhering to the inside of the culture tank due to the growth of bacteria, bubble column culture tanks are not equipped with stirring devices such as stirring rods or stirring blades inside the culture tank; It is equipped with an oxygen-containing gas introduction mechanism for the purpose of supplying M and stirring the contents during culture. The upper part of the culture tank is provided with an outlet for discharging the introduced gas, and the bottom part is provided with an outlet for taking out the culture solution.
又、培養槽の解体洗浄をｌ！ｉ単に実施する為、その底
部もしくは包泡塔培薔槽の壁面に開放機構を具備してい
る。Also, disassemble and clean the culture tank! i For simple implementation, an opening mechanism is provided at the bottom or on the wall of the bubble wrapper cultivation tank.
酸素含有ガスの導入機構は、気泡塔培養槽の下部又は底
部の壁面に直接該ガスの吹出孔を少なくとも一つ有する
か、又は気泡塔培養槽の下部の壁面より内部に突出した
部分に少なくとも一つの吹出孔を有するものであれば良
い。槽内部に突出した構造の場合は培養時に付着したゲ
ルを容易に洗浄できる構造であることが好ましい。The introduction mechanism for the oxygen-containing gas has at least one blow-off hole for the gas directly on the wall surface of the lower part or bottom of the bubble column culture tank, or has at least one blow-off hole for the gas directly on the wall surface of the lower part of the bubble column culture tank. It is sufficient if it has two blow-off holes. In the case of a structure that protrudes into the inside of the tank, it is preferable that the structure is such that gel attached during culture can be easily washed away.
上述の気泡塔培養槽に萌述の培地を張込み、１１０〜１
２０℃で１０〜３０分間殺菌処理した後、シードを０．
１〜５０％、好ましくは０．５〜１０％接種し、０．０
１〜２ Ｖ、Ｖ、Ｉｌ 、好ましくは０．１〜０．５
Ｖ、Ｖ、ｍの割合でｍ県含有ガスを通気し２０〜３５℃
、好ましくは２５〜３３℃で２４〜１２０時間、好まし
くは２４〜７２時間培養する。The above-mentioned bubble column culture tank was filled with Moe's medium, and 110-1
After sterilization at 20°C for 10-30 minutes, the seeds were sterilized at 0.
1-50%, preferably 0.5-10% inoculation, 0.0%
1-2 V, V, Il, preferably 0.1-0.5
Aerate the gas containing m at the ratio of V, V, m and heat it to 20-35℃
, preferably at 25 to 33°C for 24 to 120 hours, preferably 24 to 72 hours.
ここで通気する酸素含有ガスとしては、酸素ガス、Ｒ素
と窒素の混合ガス、殺菌処理された空気等を例示し得る
。Examples of the oxygen-containing gas to be vented here include oxygen gas, a mixed gas of R element and nitrogen, and sterilized air.
気泡塔培養槽内部には撹拌羽根がなく、導入する酸素含
有ガスのみによって液の混合を行なうために、酸素含有
ガスの通気量が０．０１ｖ、ｖ、ｎ＋未満であると、気
泡塔培養槽の培地の混合が不充分であり且つ、酸木の供
給が不充分となり好ましくない。There is no stirring blade inside the bubble column culture tank, and the liquid is mixed only by the introduced oxygen-containing gas. Therefore, if the aeration rate of oxygen-containing gas is less than 0.01v, v, n+, the bubble column culture tank This is not preferable because the mixing of the medium is insufficient and the supply of acid wood is insufficient.
また、酸素含有ガスの通気量が２Ｖ、 Ｖ、 ＋１を越
えると培地の撹拌が激しくなりすぎて好ましくない。Furthermore, if the amount of oxygen-containing gas aerated exceeds 2 V, V, +1, stirring of the culture medium becomes too vigorous, which is not preferable.
本発明の棚段静置培養器は１２０℃で変形しない耐熱性
樹脂、例えばポリカーボネート製又は金属製トレーを棚
枠上に段状に積み重ねた培養器である。各トレーは、そ
の培養液表面を酸素含有ガスが０〜５０００１１１７分
、好ましくは０，２〜２０００ｔｅ ／分の割合で通気
し得るように隙間がおいて積み重ねられている。或いは
、酸素含有ガスが上述の割合で通気するように上部側面
に通気孔のおいているトレーを積み重ねても良い。棚段
状に積み重ねられたトレーは、酸素含有ガス導入機構と
排出機構を具備するボックスの中に収納される。The plated stationary incubator of the present invention is an incubator in which trays made of a heat-resistant resin that does not deform at 120° C., such as polycarbonate or metal, are stacked in tiers on a shelf frame. Each tray is stacked with gaps such that oxygen-containing gas can be passed through the surface of the culture solution at a rate of 0 to 50001117 minutes, preferably 0.2 to 2000 te/min. Alternatively, trays may be stacked with ventilation holes in the upper sides so that the oxygen-containing gas is vented at the rate described above. The trays stacked in tiers are housed in a box equipped with an oxygen-containing gas introduction mechanism and an exhaust mechanism.
本発明における静置培養は、殺菌処理された上述の棚段
静置培養器の各トレーに気泡塔培養された培養液を分注
し、トレーの培養液表面を酸素含有ガスがθ〜５０００
ａ！ ／分の割合で通気するように該ガスを導入し、２
０〜３５℃で３〜２５日門静置培養する。In the static culture in the present invention, the culture solution cultured in a bubble column is dispensed onto each tray of the above-mentioned plated static culture device that has been sterilized, and the surface of the culture solution on the tray is heated to an oxygen-containing gas of θ ~ 5000.
a! The gas was introduced so as to aerate at a rate of 2 minutes.
Static culture is performed for 3 to 25 days at 0 to 35°C.
棚段静置培養器の殺菌処理は温度１１０〜１２０℃で０
．３〜２時間、好ましくは１０〜１００ｄ／ｊ！の割合
で水を存在させながらおこなわれる。Sterilization treatment of shelf static culture vessels is carried out at a temperature of 110 to 120℃.
．． 3-2 hours, preferably 10-100 d/j! It is carried out in the presence of water at a ratio of
（発明の効果）
本発明の方法は、微生物セルロースの大間生産に適して
おり、短かい培養日数、且つ高い収率で、微生物セルロ
ースを得ることが出来る。(Effects of the Invention) The method of the present invention is suitable for the production of microbial cellulose, and microbial cellulose can be obtained in a short culture period and at a high yield.
例えば、シード培養工程とコンテノーにょる静置培養工
程とからなる従来の培養方法に比べて、本発明の方法は
およそ２／３のＰｉ ｎ日数で、少なくとも２０％増の
収率を得ることが出来る。For example, compared to a conventional culture method consisting of a seed culture step and a static container culture step, the method of the present invention can obtain at least a 20% increase in yield in approximately two-thirds the Pin number of days. I can do it.
更に、本発明の方法によれば、気泡塔培養槽の洗浄、静
置１ａ！器の組立及び洗浄等において大巾な時間短縮及
び労力の省力化を達成することが出来る。Furthermore, according to the method of the present invention, the bubble column culture tank is cleaned and left standing 1a! Significant time and labor savings can be achieved in the assembly and cleaning of vessels.
以下、実流例によって本発明を具体的に説明するが、本
発明はこれら実施例に限定されるものではない。Hereinafter, the present invention will be specifically explained using actual flow examples, but the present invention is not limited to these examples.
１五１
内容量５００ｍの肩付振盪フラスコに、蔗糖５ｇ／旧、
イーストエクストラクト０．５ｇ／ｄｌ、（ＮＨ４）２
Ｓ０４０．５ｇ／旧、Ｋ１−１，２ ｐｏ４０．３ｇ／
ｄｌ、Ｍ（Ｊ ＳＯ７１−１２０ｏ、ｏ５ｇ、／旧ＴＭ
Ａ−８２１（東芝シリコン社製’） ０．００１ｄ／
ｄｌを含有する培地（１））Ｉ＝５．０ ）を４００ａ
ｉｔ分注し、Ａ−トクレーブで１２０℃、２０分間殺菌
処理し、予めリフレッシ−したＡ ｃｅｔｏｂａｃｔｏ
ｒ ａｃｅ口５ｕｂｓｐ。151 Into a shoulder shaker flask with a capacity of 500 m, add 5 g of sucrose/old,
Yeast extract 0.5g/dl, (NH4)2
S040.5g/old, K1-1,2 po40.3g/
dl, M (J SO71-120o, o5g, / former TM
A-821 (manufactured by Toshiba Silicon Corporation) 0.001d/
dl-containing medium (1))I=5.0) at 400a
It was dispensed and sterilized in an A-toclave at 120°C for 20 minutes, and refreshed in advance.
r ace mouth 5 ubsp.
ｘｙｌｉｎｕｍ ＡＴＣＣ１０８２１菌株を接種し、振
盪機上＜１０１００ｒｐ 、３０℃で４８時間培養した
。xylinum ATCC10821 strain and cultured for 48 hours at 30°C on a shaker at <10,100 rp.
培養槽内底部に酸素含有ガス導入機構を具備し、培養槽
上部には導入ガス刊出口及びその底部には培晶液取出口
が設けられており、底部が開放されるようになっている
０１１７５−の内容量の気泡塔培養槽にシード培養と同
じ培地（ｐＨ＝４）を１５０！張込み１２０℃で２０分
間バッチ殺菌をした。The bottom of the culture tank is equipped with an oxygen-containing gas introduction mechanism, the top of the culture tank is equipped with an inlet gas outlet, and the bottom of the tank is provided with a culture liquid outlet, so that the bottom is open.01175 - Add the same medium (pH = 4) as for seed culture to a bubble column culture tank with an internal capacity of 150! Batch sterilization was carried out at 120° C. for 20 minutes.
殺菌処理後、シード培養で得られたシード液（菌数５．
７Ｘ １０７個／ｄ） １％を接種し、殺菌処理され
た空気を０．２５ｖ、ｖ」の割合で通気しながら３０℃
で７２時間培養した（培養６９時間後の菌数は１×１０
８個／ｄであった）。After sterilization, the seed liquid obtained by seed culture (the number of bacteria is 5.
(7
Cultured for 72 hours (the number of bacteria after 69 hours of culture was 1 x 10
8 pieces/d).
オートクレーブで１２０℃、６０分間殺菌処理された棚
段静置培養器の各々のトレー（ｌｌｌＸ横×高さ＝ ５
３０ｍ Ｘ ４２０ｓ＋ Ｘ ８０Ｍ）に気泡塔培養液
を４１分注した。棚段静置培養器のトレーの数は９枚で
ある。殺菌された空気を棚段静置培養器のトレー内の培
地表面の通気割合が１２／分となるように導入し、３１
℃、１０日間培落した。Each tray of a shelf static incubator was sterilized in an autoclave at 120°C for 60 minutes (ll x width x height = 5
The bubble column culture solution was dispensed into 41 portions (30 m x 420s + x 80M). The number of trays in the plated static incubator is nine. Sterilized air was introduced so that the ventilation rate of the culture medium surface in the tray of the plated static incubator was 12/min.
The cells were cultured at ℃ for 10 days.
培養１０日後のトレー当りのゲル量は３．１２Ｋｇ、ゲ
ル厚は１１〜１５麺、収率（重量／張込液ｍ）は７８．
０％で、棚段塔ｉ器当りのゲル量は２８．０８ Ｋｇで
あった。After 10 days of culture, the amount of gel per tray was 3.12 kg, the gel thickness was 11 to 15 noodles, and the yield (weight/m of filling solution) was 78.
At 0%, the amount of gel per tray column was 28.08 Kg.
気泡塔培養槽の洗浄、静置培養器の洗浄、培養液の分注
及び培養状態の観察等の操作が容易であり、大巾な処理
時間の短縮及び労力の省力化がなされた。Operations such as cleaning the bubble column culture tank, cleaning the static culture vessel, dispensing the culture solution, and observing the culture state are easy, and the processing time and labor are greatly reduced.
Ｌ較旦
実施例で得られたシードを各々のコンテナー（縦×横Ｘ
高さ＝６４５麟×３８５朧×１５０履）に張込まれてい
る実施例と同じ培地（５１）に接種した。The seeds obtained in the L comparison example were placed in each container (length x width
The same medium (51) as in the example was inoculated into a medium (height = 645 cm x 385 cm x 150 cm).
殺晶された空気を当該培地表面の通気紙が１５０ＩｄＺ
分となるように通気し、３１．５℃で２０日間培養した
。培養２２日後のコンテナー当りのゲル輩ｔよ３．１に
９、ゲル厚さは１０〜１３Ｍ、収率（重石／張込液ＦＭ
）は６２％であった。The aeration paper on the surface of the culture medium transports the crystallized air to 150IdZ.
The cells were aerated for 20 minutes and cultured at 31.5°C for 20 days. After 22 days of culture, the gel mass per container was 3.1 to 9, the gel thickness was 10 to 13 M, and the yield (weight/filling solution FM
) was 62%.
喀Ｘ１２キボｆ匈 越 正丸喀
Claims (4)
ｘｙｌｉｎｕｍをシード培養し、攪拌機を具備していな
い気泡塔培養槽中の培地にシードを接種して、酸素含有
ガスを通気しながら気泡塔培養し、 得られた培養液を棚段培養器に分注して、静置培養する
ことから成る微生物セルロースの産生方法。(1) Acetobacteracetisubsp.
xylinum, inoculate the seeds into a medium in a bubble column culture tank without a stirrer, culture in a bubble column while aerating oxygen-containing gas, and divide the obtained culture solution into tray culture vessels. A method for producing microbial cellulose, which comprises injecting the cellulose and culturing it statically.
ラクト又は総合アミノ酸及びフィチン酸；（ＮＨ＿４）
＿２ＳＯ＿４；ＫＨ＿２ＰＯ＿４；及びＭｇＳＯ＿４を
含有する培地で２０〜３５℃、２４〜９６時間を行うこ
とから成る特許請求の範囲第１項に記載の方法。(2) Seed culture with sucrose or sugar; yeast extract or general amino acids and phytic acid; (NH_4)
2. The method according to claim 1, comprising carrying out the incubation at 20-35° C. for 24-96 hours in a medium containing _2SO_4; KH_2PO_4; and MgSO_4.
ラクト又は総合アミノ酸及びフィチン酸；（ＮＨ＿４）
＿２ＳＯ＿４；ＫＨ＿２ＰＯ＿４；及びＭｇＳＯ＿４を
含有する培地に０．１〜５０％接種して酸素含有ガスを
０．０１〜２ｖ．ｖ．ｍの割合で通気しながら２０〜３
５℃、２４〜１２０時間行なうことから成る特許請求の
範囲第１項に記載の方法。(3) Bubble column culture with sucrose or sugar; yeast extract or general amino acids and phytic acid; (NH_4)
A medium containing _2SO_4; KH_2PO_4; and MgSO_4 was inoculated at 0.1-50%, and oxygen-containing gas was added at 0.01-2v. v. While ventilating at a rate of 20 to 3 m
2. A method according to claim 1, comprising carrying out at 5 DEG C. for 24 to 120 hours.
分の割合で培養液表面に通気しながら、２０〜３５℃、
１２〜６００時間行なうことから成る特許請求の範囲第
１項に記載の方法。(4) Static culture with 0 to 5000 ml/oxygen-containing gas
20-35℃, while aerating the culture solution surface at a rate of 20-35℃.
2. A method according to claim 1, comprising carrying out for 12 to 600 hours.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63071071A JP2797308B2 (en) | 1988-03-25 | 1988-03-25 | Method for producing microbial cellulose |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP63071071A JP2797308B2 (en) | 1988-03-25 | 1988-03-25 | Method for producing microbial cellulose |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH01243997A true JPH01243997A (en) | 1989-09-28 |
| JP2797308B2 JP2797308B2 (en) | 1998-09-17 |
Family
ID=13449925
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP63071071A Expired - Fee Related JP2797308B2 (en) | 1988-03-25 | 1988-03-25 | Method for producing microbial cellulose |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2797308B2 (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5079162A (en) * | 1986-08-28 | 1992-01-07 | Weyerhaeuser Company | Reticulated cellulose and methods and microorganisms for the production thereof |
| US5144021A (en) * | 1985-10-18 | 1992-09-01 | Weyerhaeuser Company | Reticulated cellulose and methods and microorganisms for the production thereof |
| US5821109A (en) * | 1985-10-18 | 1998-10-13 | Monsanto Life Sciences Co. | Reticulated cellulose and methods and microorganisms for the production thereof |
| US5871978A (en) * | 1985-10-18 | 1999-02-16 | Monsanto Life Sciences Co | Method of producing reticulated cellulose having type II crystalline cellulose |
| US6979450B2 (en) | 1998-02-06 | 2005-12-27 | Japan Bcg Laboratory | Method and compositions for detection and diagnosis of infectious diseases |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6287099A (en) * | 1985-10-14 | 1987-04-21 | Ajinomoto Co Inc | Production of cellulosic substance by bacterium |
-
1988
- 1988-03-25 JP JP63071071A patent/JP2797308B2/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6287099A (en) * | 1985-10-14 | 1987-04-21 | Ajinomoto Co Inc | Production of cellulosic substance by bacterium |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5144021A (en) * | 1985-10-18 | 1992-09-01 | Weyerhaeuser Company | Reticulated cellulose and methods and microorganisms for the production thereof |
| US5821109A (en) * | 1985-10-18 | 1998-10-13 | Monsanto Life Sciences Co. | Reticulated cellulose and methods and microorganisms for the production thereof |
| US5871978A (en) * | 1985-10-18 | 1999-02-16 | Monsanto Life Sciences Co | Method of producing reticulated cellulose having type II crystalline cellulose |
| US6329192B1 (en) | 1985-10-18 | 2001-12-11 | Cp Kelco U.S., Inc. | Reticulated cellulose and methods of microorganisms for the production thereof |
| US6429002B1 (en) | 1985-10-18 | 2002-08-06 | Cp Kelco U.S., Inc. | Reticulated cellulose producing acetobacter strains |
| US5079162A (en) * | 1986-08-28 | 1992-01-07 | Weyerhaeuser Company | Reticulated cellulose and methods and microorganisms for the production thereof |
| US6979450B2 (en) | 1998-02-06 | 2005-12-27 | Japan Bcg Laboratory | Method and compositions for detection and diagnosis of infectious diseases |
| US7655218B2 (en) | 1998-02-06 | 2010-02-02 | Japan Bcg Laboratory | Methods and compositions for detection and diagnosis of infectious diseases |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2797308B2 (en) | 1998-09-17 |
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