JP3433300B2 - Highly producing yeast cell wall lytic enzyme and method for lysing yeast cell wall using the same - Google Patents
Highly producing yeast cell wall lytic enzyme and method for lysing yeast cell wall using the sameInfo
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- JP3433300B2 JP3433300B2 JP21108193A JP21108193A JP3433300B2 JP 3433300 B2 JP3433300 B2 JP 3433300B2 JP 21108193 A JP21108193 A JP 21108193A JP 21108193 A JP21108193 A JP 21108193A JP 3433300 B2 JP3433300 B2 JP 3433300B2
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- cell wall
- negative
- yeast cell
- enzyme
- bacterium
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Description
【０００１】[0001]
【産業上の利用分野】本発明は、オエルスコフィア（Oe
rskovia) に属する酵母細胞壁溶解酵素（YLase)を著量
生産する能力を有するＥ24株（菌寄第13692 号）の生産
する酵素または培養液を用いて酵母細胞壁を溶解する方
法、さらにはこの酵素または培養液を用いて酵母細胞壁
を溶解することを特徴とする酵母エキスを製造する方法
に関する。BACKGROUND OF THE INVENTION The present invention is applicable to the Oerskophia ( Oe
rskovia ) yeast cell wall lysing enzyme (YLase), which has the ability to produce a large amount of yeast cell wall lysing enzyme using the enzyme or culture medium produced by the E24 strain (No. 13692). The present invention relates to a method for producing a yeast extract, which comprises dissolving a yeast cell wall using a culture solution.
【０００２】[0002]
【従来の技術】酵母エキスの製造は、主に自己溶解させ
る方法で行われているが、最近は、呈味性の高い酵母エ
キス製造方法として、市販の酵母溶解酵素（YLase)剤を
用いた酵素溶解法もよく用いられるようになっている。
この酵母溶解酵素(YLase) を生産する微生物は、これま
でにアルスロバクター（Arthrobacter) 属 (特公昭47-3
2674、特公昭48-2790)、オエルスコフィア（Oerskovia)
属（特公昭52-46308、特公昭63-51676) 、ストレプトミ
セス（Streptomyces) 属（特公昭52-31427、特公昭60-2
2916) など多くの菌種が報告されており、すでに酵素剤
として数種市販されている（工業用酵素として、YL-15
(天野製薬株式会社) 、ツニカーゼ（大和化成株式会
社）等。研究用試薬として、ザイモリエイス（生化学工
業株式会社）、キタラーゼ（和光純薬工業株式会社）
等）。BACKGROUND ART Yeast extract is mainly produced by a method of self-dissolving, but recently, as a method for producing a yeast extract having a high taste, a commercially available yeast lysing enzyme (YLase) agent has been used. Enzyme-dissolving methods are also becoming popular.
This yeast lytic enzyme (YLase) -producing microorganism has been the genus of Arthrobacter (Article 47-3).
2674, JP-B-48-2790), Oerusukofia (Oerskovia)
Genus (JP-B-52-46308, JP-B-63-51676), Streptomyces (Streptomyces) belonging to the genus (JP-B-52-31427, JP-B-60-2
2916) and many other bacterial species have been reported, and several of them have already been marketed as enzymatic agents (YL-15
(Amano Pharmaceutical Co., Ltd.), Tunicase (Daiwa Kasei Co., Ltd.), etc. As research reagents, Zymolyce (Seikagaku Corporation), Kitarase (Wako Pure Chemical Industries, Ltd.)
etc).
【０００３】[0003]
【発明が解決しようとする課題】しかし、工業用として
市販されている酵素は活性が不充分であり、酵母の溶解
に多量（酵母乾燥重量に対して酵素剤１〜数％）の酵素
剤を必要とする。このため、酵母エキスの製造コストに
占める酵素剤の費用が多くなる。一方、研究用試薬とし
て販売されている酵素は、活性が高いが高価である。However, the enzyme commercially available for industrial use has insufficient activity, and a large amount (1 to several% of the enzyme preparation based on the dry weight of the yeast) of the enzyme preparation is used to dissolve the yeast. I need. Therefore, the cost of the enzyme preparation in the production cost of the yeast extract increases. On the other hand, the enzyme sold as a research reagent has high activity but is expensive.
【０００４】また、前記の微生物による酵母溶解酵素(Y
Lase) 生産においては、培地成分による酵素生産の抑制
が報告されており（Andrews, B. A. and Asenjo J. A.:
Biotechnol. Bioeng., 30, 628-637 (1987)) 、通常の
バッチ培養では菌体増殖が進んだ増殖後期において培地
成分が少なくなってから酵素を生産する（Kitamura,K.,
Kaneko, T. and Yamamoto, Y. : J. Gen. Appl. Micro
biol., 18, 57-71 (1972)) 。従って、培養液の酵素活
性が高くならず、酵素の効率的な生産に不利であり、酵
素活性を高めるために、炭素源を制限した連続培養など
の方法がとられることがある。In addition, the yeast lytic enzyme (Y
In Lase) production, suppression of enzyme production by medium components has been reported (Andrews, BA and Asenjo JA:
Biotechnol. Bioeng., 30 , 628-637 (1987)), an enzyme is produced after the number of medium components is reduced in the latter stage of growth in which cell growth has progressed in normal batch culture (Kitamura, K.,
Kaneko, T. and Yamamoto, Y .: J. Gen. Appl. Micro
biol., 18 , 57-71 (1972)). Therefore, the enzyme activity of the culture broth does not increase, which is disadvantageous to the efficient production of the enzyme, and in order to increase the enzyme activity, a method such as continuous culturing with a limited carbon source may be adopted.
【０００５】かかる問題を解決するために、本発明者ら
は酵母溶解能を有する微生物を広く天然界より検索し、
排水処理場近傍の土壌より分離したＥ24が酵母細胞壁を
強力に溶解し、また炭素源及び窒素源を多く含有する培
地でも酵素生産が抑制されることがなく、菌の増殖に伴
って酵母細胞壁溶解酵素を生産することを見出し、本発
明を完成した。In order to solve such a problem, the present inventors extensively searched for microorganisms having the ability to dissolve yeast in the natural world,
Culture of E24 isolated from wastewater treatment plant near the soil strongly loosened dissolved the yeast cell walls, also containing much carbon and nitrogen sources
Enzyme production is not suppressed even on the ground,
As a result, they found that yeast cell wall lysing enzyme was produced, and completed the present invention.
【０００６】[0006]
【課題を解決するための手段】本発明は、酵母細胞壁溶
解酵素を著量生産する能力を有するオエルスコフィア
（Oerskovia sp.) Ｅ24 (工業技術院生命工学工業研究
所菌寄託番号第13692 号)である。また、本発明は、前
記菌または該菌が生産する酵母細胞壁溶解酵素を使用す
ることを特徴とする酵母細胞壁の溶解方法である。さら
に本発明は、前記の菌または該菌が生産する酵母細胞壁
溶解酵素を使用することを特徴とする酵母エキスの製造
方法である。The present invention is Oerskovia sp. E24 (Deposit No. 13692, Institute of Biotechnology, Institute of Industrial Science and Technology), which has the ability to produce yeast cell wall lysing enzyme in a large amount. . Further, the present invention is a method for lysing a yeast cell wall, which comprises using the bacterium or a yeast cell wall lysing enzyme produced by the bacterium. Furthermore, the present invention is a method for producing a yeast extract, characterized by using the above-mentioned bacterium or a yeast cell wall lysing enzyme produced by the bacterium.
【０００７】本発明のオエルスコフィア（Oerskovia s
p.) Ｅ24は、寄託番号第13692 号にて工業技術院生命工
学工業研究所に寄託されている。本菌の特性を列挙すれ
ば次の通りである。
Ｉ．形態及び培養的性質（肉汁寒天培地 30℃)
１．形態：培養初期は、長桿菌であり一部分枝した形態
が見られる。培養後期に至ると単桿菌から球菌に近い形
態を示す。
２．内生胞子：形成しない。
３．運動性：認められない。
４．鞭毛：認められない。
５．グラム染色：陽性。
６．コロニー：隆起している。
７．色素：培養後期に黄色色素を産生する。The Oerskovia s of the present invention
p.) E24 has been deposited at the Institute of Biotechnology, Institute of Industrial Science, under the deposit number 13692. The characteristics of this bacterium are listed below. I. Morphology and cultural properties (meat agar medium 30 ° C) 1. Morphology: In the early stage of culture, it is a long-rod bacterium and a partially branched morphology is seen. In the latter half of the culture, it shows a morphology similar to monococcus to cocci. 2. Endospores: Do not form. 3. Motility: Not recognized. 4. Flagella: Not recognized. 5. Gram stain: positive. 6. Colony: Raised. 7. Pigment: A yellow pigment is produced in the latter stage of culture.
【０００８】II．生理的性質
１．生育条件：20〜37℃の温度範囲で生育でき、42℃で
は生育しない。また、嫌気条件下でも生育する。
２．硫化水素生成：陰性。
３．インドール生成：陰性。
４．カタラーゼ：陽性。
５．オキシダーゼ：陰性。
６．硝酸塩還元性：陽性であるがＮ2までは還元しな
い。
７．ゼラチン液化：陽性。II. Physiological properties 1. Growth conditions: It can grow in the temperature range of 20-37 ℃, but not at 42 ℃. It also grows under anaerobic conditions. 2. Hydrogen sulfide production: Negative. 3. Indole formation: negative. 4. Catalase: positive. 5. Oxidase: negative. 6. Nitrate reduction: Positive but does not reduce to N 2 . 7. Gelatin liquefaction: Positive.
【０００９】８．ウレアーゼ：陰性。
９．アルギニンジヒドロラーゼ：陰性。
10．リジンデカルボキシラーゼ：陰性。
11．オルニチンデカルボキシラーゼ：陰性。
12．トリプトファンデアミナーゼ：陰性。
13．β−ガラクトシダーゼ：陽性。
14．糖類の資化性：グルコース、アラビノース、マンノ
ース、Ｎ−アセチル−Ｄ−グルコサミン、マルトースを
資化し、マンニトールを資化しない。8. Urease: Negative. 9. Arginine dihydrolase: negative. Ten. Lysine decarboxylase: negative. 11. Ornithine decarboxylase: negative. 12. Tryptophan deaminase: negative. 13. β-galactosidase: positive. 14. Utilization of sugars: Utilizes glucose, arabinose, mannose, N-acetyl-D-glucosamine and maltose, but does not utilize mannitol.
【００１０】以上の菌学的知見から本菌はオエルスコフ
ィア（Oerskovia）属に属するものであるが、既に報告
されている同属のオエルスコフィア・ツルバータ（Oers
kovia turbata）、オエルスコフィア・キサンチネオリ
チカ（Oerskovia xanthineolytica)、オエルスコフィア
・シトリア（Oerskovia citrea) とは一部の性質、例え
ば鞭毛の有無、運動性などの点が異なっている。[0010] The above, but this bacteria from mycological findings are those belonging to the Oerusukofia (Oerskovia) genus, of the same genus that have already been reported Oerusukofia-Tsurubata (Oers
It differs from kovia turbata ), Oerskovia xanthineolytica ( Oerskovia xanthineolytica) and Oerskovia citrea ( Oerskovia citrea ) in some properties such as flagella presence and motility.
【００１１】本発明ではオエルスコフィア（Oerskovi
a）Ｅ24株を液体培地または固体培地で培養するが、酵
母細胞壁溶解酵素を取得するには液体培地の方が便利で
ある。培地としては、用いられる菌が資化できる炭素
源、窒素源、無機塩類、その他微量栄養源を適当な濃度
で含むことが望ましい。本菌は炭素源及び窒素源を多く
含有する培地においても酵素生産が抑制されることな
く、菌の増殖に伴って酵母細胞壁溶解酵素を生産するの
で、培地としてビール工場の余剰酵母や酵母エキス製造
における残渣（ＹＣＷ）をそのまま利用することがで
き、安価に酵素を生産できる。酵素は、通常の酵素の精
製法に従って精製して使用することができるし、培養上
清をそのまま用いても良い。In the present invention, the Oerskophia (Oerskovi
a) E24 strain is cultivated in liquid medium or solid medium,
Liquid medium is more convenient for obtaining mother cell wall lysing enzyme
is there. As a medium, carbon that can be assimilated by the bacteria used
Sources, nitrogen sources, inorganic salts, and other trace nutrient sources in appropriate concentrations
It is desirable to include in. This bacterium has many carbon and nitrogen sources
Enzyme production is not suppressed even in the medium containing
Production of yeast cell wall lysing enzymes as the bacteria grow
In the production of surplus yeast and yeast extract in the brewery as a medium
The residue (YCW) in can be used as it is.
The enzyme can be produced inexpensively. Enzyme is a normal enzyme
It can be purified according to the manufacturing method and used.
You may use Kiyoshi as it is.
【００１２】本発明に使用する酵母細胞壁溶解酵素は、
45℃まで、またpH５〜９の間で安定である。反応至適温
度は40〜50℃、至適pHは６〜９と広い範囲で有効であ
る。酵母細胞壁分解酵素活性は、キタムラら（Kitamur
a, K., Kaneko, T. and Yamamoto, Y. : J. Gen. Appl.
Microbiol., 18, 57-71 (1972)) が用いた酵母溶解活
性測定方法を一部次のように改変して測定した。基質
は、ＹＣＷ溶液（吸光度(800nm) 約 1.0、pH 7.5) を用
い、緩衝液は、pH 7.5の1/15Ｍリン酸緩衝液を用いた。
反応は、ＹＣＷ溶液３mlに緩衝液１mlを加え、37℃に保
温した後に酵素液１mlを加えて行った。この反応液の吸
光度（800nm)を酵素液添加直後（吸光度Ｂ）および30分
または１時間後（吸光度Ａ）に測定し、次式から酵素活
性を算出した。The yeast cell wall lysing enzyme used in the present invention is
It is stable up to 45 ° C and between pH 5-9. The optimum reaction temperature is 40 to 50 ° C., and the optimum pH is 6 to 9, and it is effective in a wide range. Yeast cell wall degrading enzyme activity was determined by Kitamura et al.
a, K., Kaneko, T. and Yamamoto, Y .: J. Gen. Appl.
Microbiol., 18 , 57-71 (1972)) was used, and the method for measuring the yeast lytic activity was partially modified as follows. A YCW solution (absorbance (800 nm) about 1.0, pH 7.5) was used as a substrate, and a 1/15 M phosphate buffer having a pH of 7.5 was used as a buffer.
The reaction was carried out by adding 1 ml of buffer solution to 3 ml of YCW solution, keeping the temperature at 37 ° C., and then adding 1 ml of enzyme solution. The absorbance (800 nm) of this reaction solution was measured immediately after the addition of the enzyme solution (absorbance B) and after 30 minutes or 1 hour (absorbance A), and the enzyme activity was calculated from the following formula.
【００１３】[0013]
【数１】 [Equation 1]
【００１４】酵素の生産は、得られた変異株（Ｅ24株）
及び、対照としてYLase 生産菌オエルスコフィア・キサ
ンチネオリチカ（Oerskovia xanthineolytica JCM3164)
について行った。各菌株の生産する酵素を市販酵素剤と
比較するために、−80℃アセトン沈澱法により各菌株培
養液から粗酵素を調製し、その酵素活性および蛋白質当
たりの比活性を比較した。結果を下表に示し、本菌の優
位性を示す。
培養上清および粗酵素の活性
菌株名または酵素名 YLase
培養液上清〔U/ml〕粗酵素剤〔U/mg〕比活性〔U/mg蛋白〕
Ｅ24 1220 440 4630
JCM3164 62 19 380
YL-15 - 18 300
ザイモリエイス - 960 3430 The enzyme was produced by the obtained mutant strain (E24 strain).
And as a control, YLase-producing bacterium Oerskovia xanthineolytica JCM3164
I went about. In order to compare the enzyme produced by each strain with a commercially available enzyme preparation, a crude enzyme was prepared from the culture solution of each strain by the -80 ° C acetone precipitation method, and the enzyme activity and the specific activity per protein were compared. The results are shown in the table below, showing the superiority of this bacterium. Activity of culture supernatant and crude enzyme Strain name or enzyme name YLase culture solution supernatant [U / ml] Crude enzyme agent [U / mg] Specific activity [U / mg protein] E24 1220 440 4630 JCM3164 62 19 380 YL-15 -18 300 Zymory Ace-960 3430
【００１５】[0015]
【実施例】以下、実施例により本発明を具体的に説明す
る。
実施例１
本菌の分離は、ＹＣＷ 1％、グルコース 0.5％、酵母エ
キス 1％、寒天 1.5％、pH7.5 からなる培地上に採取し
た土壌懸濁液を適宜希釈して塗布し、37℃で培養した。
コロニー周辺に大きなハローを形成する菌はグルコース
及び酵母エキスによる酵素生産抑制を受けず、酵素生産
量が多い菌と判断し、ハローの大きなコロニーを分離
し、Ｅ24と命名した。EXAMPLES The present invention will be specifically described below with reference to examples. Example 1 Isolation of this bacterium was carried out by appropriately diluting the soil suspension collected on a medium consisting of YCW 1%, glucose 0.5%, yeast extract 1%, agar 1.5%, pH 7.5, and applying it at 37 ° C. It was cultured in.
A bacterium that forms a large halo around the colony was judged to be a bacterium having a large amount of enzyme production without being inhibited by the glucose and yeast extract, and a large halo colony was isolated and named E24.
【００１６】本菌を用いる酵母細胞壁溶解酵素生産は、
培地にＹＣＷを使用して振盪フラスコまたはジャーファ
ーメンターを用いて実施した。フラスコ培養は、培地と
して１％ＹＣＷ 100mlを 500ml容振盪フラスコに入れ、
37℃で３日間培養した。培養終了後、遠心分離（10,000
ｇ、15分) にて培養上清を取得し、その酵母溶解活性を
測定したところ、1220U/mlであった。酵素活性の測定は
前述した方法を用いて行った。Yeast cell wall lysing enzyme production using this bacterium is
Performed using shake flasks or jar fermenters with YCW as the medium. The flask culture was carried out by placing 100 ml of 1% YCW in a 500 ml shake flask as a medium,
It was cultured at 37 ° C for 3 days. After culturing, centrifuge (10,000
(g, 15 minutes), the culture supernatant was obtained, and its yeast lytic activity was measured to be 1220 U / ml. The enzyme activity was measured using the method described above.
【００１７】一方、ジャーファーメンターを用いた酵素
生産の条件は以下の通りであった。 項目 設定
使用ジャーファーメンター：丸菱バイオエンジ (株) 製 MDL-500
培地 ：５％ＹＣＷ、pH 7.5、２Ｌ
温度 ：30℃
攪拌数 ：300rpm
通気量 ：0.5 vvm pH調整 ：培養中 NaOHで 7.5に調整
図１に培養中の酵素活性の経時変化を示す。得られた培
養液から遠心分離にて菌体を除去し、粗酵素液とした。On the other hand, an enzyme using a jar fermenter
The production conditions were as follows. Item settings
Jar fermenter used: MDL-500 manufactured by Maruhishi Bioengineering Co., Ltd.
Medium: 5% YCW, pH 7.5, 2L
Temperature: 30 ℃
Stirring number: 300 rpm
Airflow: 0.5 vvmpH adjustment: Adjusted to 7.5 with NaOH during culture
Figure 1 shows the time course of enzyme activity during culture. Obtained culture
The bacterial cells were removed from the nutrient solution by centrifugation to obtain a crude enzyme solution.
【００１８】実施例２
実施例１で得られた粗酵素液を用い乾燥酵母スラリーを
原料として酵母エキス製造を実施した。乾燥酵母スラリ
ー（乾燥酵母10％）に培養液をスラリー 100ml当たり１
ml添加し、45℃、17時間、緩やかな攪拌条件下で酵母エ
キス製造を行った。比較として酵母エキス製造に用いら
れている市販酵素YL-15(天野製薬株式会社製) を固形物
換算 1.0％（ｗ／ｗ）になるように加えて同じ条件下で
酵母エキス製造を行った。反応終了後、遠心分離（8000
ｇ、10分）にて上清と沈澱に分離し、それぞれの乾燥重
量を測定した。エキス収量は、次のように定義して比較
した。Example 2 Using the crude enzyme solution obtained in Example 1, yeast extract was produced using a dry yeast slurry as a raw material. 1 per 100 ml slurry of dry yeast slurry (dry yeast 10%)
ml was added, and yeast extract was produced at 45 ° C. for 17 hours under mild stirring conditions. For comparison, a commercially available enzyme YL-15 (manufactured by Amano Pharmaceutical Co., Ltd.) used for yeast extract production was added so as to be 1.0% (w / w) in terms of solid matter, and yeast extract was produced under the same conditions. After completion of the reaction, centrifuge (8000
(g, 10 minutes), the supernatant and the precipitate were separated, and the dry weight of each was measured. The extract yield was defined as follows and compared.
【００１９】[0019]
【数２】 [Equation 2]
【００２０】次表にその比較を示す。
酵母エキス収量
菌株又は酵素名 添加量〔%W/W〕 エキス収量〔％〕
Ｅ24 0.3 37.6
YL-15 1.0 37.2
上表に示したように、本菌の培養液を用いた場合、活性
が高く、少量の使用でも市販酵素を使用した場合と同様
の酵母エキス収量が得られた。The following table shows the comparison. Yeast extract yield Strain or enzyme name Addition amount [% W / W] Extract yield [%] E24 0.3 37.6 YL-15 1.0 37.2 As shown in the above table, when using the culture solution of this bacterium, the activity is high, Even when used in a small amount, the yield of yeast extract was similar to that obtained when a commercially available enzyme was used.
【００２１】実施例３
Ｅ24株を用いた酵母エキスの製造を行った。乾燥酵母ス
ラリー（乾燥酵母10％）にＥ24株培養液を１％接種し、
30℃で振盪することにより酵母エキスを製造し、振盪７
日後のエキス収量を測定した。対照として、Ｅ24株を接
種せずに同様に振盪した場合のエキス収量を測定した。
エキス収量の測定方法は、実施例２と同じである。７日
振盪した場合のエキス収量の経時変化を次表に示した。
この結果から、Ｅ24株を接種することによっても、酵母
エキスを効率的に製造できることが示された。Example 3 A yeast extract was produced using the E24 strain. 1% of E24 strain culture solution was inoculated into the dry yeast slurry (dry yeast 10%),
A yeast extract was produced by shaking at 30 ° C. and shaking 7
The extract yield after day was measured. As a control, the extract yield was measured when E24 strain was similarly inoculated and not shaken.
The method for measuring the extract yield is the same as in Example 2. The following table shows the changes over time in the extract yield when shaken for 7 days.
From this result, it was shown that the yeast extract can also be efficiently produced by inoculating the E24 strain.
【００２２】 エキス収量の経時変化 エキス収量〔％〕 日数 Ｅ24接種 対照 0 12.0 -- 1 13.0 11.3 2 15.3 12.2 4 20.3 13.3 7 26.5 14.0 Change in extract yield with time Extract yield [%] Days E24 inoculation Control 0 12.0 --1 13.0 11.3 2 15.3 12.2 4 20.3 13.3 7 26.5 14.0
【００２３】[0023]
【発明の効果】本発明によれば、新規な菌オエルスコフ
ィア(Oerskovia sp.) Ｅ24は炭素源及び窒素源を多く含
有する培地においても酵素生産が抑制されることなく、
菌の増殖に伴って酵母細胞壁溶解酵素を生産するので、
培地としてビール工場の余剰酵母や酵母エキス製造にお
ける残渣（ＹＣＷ）をそのまま利用することができ、安
価に酵素を生産できる。酵素は、通常の酵素の精製法に
従って精製して使用することができるし、培養上清をそ
のまま用いることもできる。According to the present invention, a novel fungal Oerusukofia (Oerskovia sp.) E24 without enzyme production is suppressed even in a medium containing a large amount of carbon and nitrogen sources,
Since yeast cell wall lysing enzyme is produced with the growth of bacteria,
The surplus yeast in the beer factory and the residue (YCW) in the yeast extract production can be used as the medium as they are, and the enzyme can be produced at a low cost. The enzyme can be used after being purified according to a usual enzyme purification method, or the culture supernatant can be used as it is.
【図１】 実施例１において、培養中のバッチ中の酵素
活性の経時変化を示すグラフである。図中、○はYLase
〔U/ml〕、△は生菌数〔cells/ml〕を示す。FIG. 1 is a graph showing the time course of enzyme activity in a batch during culture in Example 1. In the figure, ○ is YLase
[U / ml] and Δ indicate the viable cell count [cells / ml].
───────────────────────────────────────────────────── フロントページの続き (51)Int.Cl.7 識別記号 ＦＩ (Ｃ１２Ｎ 1/20 Ｃ１２Ｎ 1/20 Ｃ１２Ｒ 1:01） Ｃ１２Ｒ 1:01 (Ｃ１２Ｎ 9/14 Ｃ１２Ｒ 1:01） Ｃ１２Ｐ 1/02 (Ｃ１２Ｐ 1/02 Ｃ１２Ｒ 1:01） (58)調査した分野(Int.Cl.7，ＤＢ名) C12N 1/00 ＢＩＯＳＩＳ／ＭＥＤＬＩＮＥ／ＷＰＩＤ Ｓ（ＳＴＮ)─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. 7 Identification code FI (C12N 1/20 C12N 1/20 C12R 1:01) C12R 1:01 (C12N 9/14 C12R 1:01) C12P 1/02 (C12P 1/02 C12R 1:01) (58) Fields investigated (Int.Cl. 7 , DB name) C12N 1/00 BIOSIS / MEDLINE / WPID S (STN)
Claims (3)
を有し、かつ下記の菌学的性質を有するオエルスコフィ
ア（Oerskovia sp.) Ｅ24 (工業技術院生命工学工業研
究所菌寄託番号第13692 号) 。Ｉ．形態及び培養的性質（肉汁寒天培地 30℃) １．形態：培養初期は、長桿菌であり一部分枝した形態
が見られる。培養後期に至ると単桿菌から球菌に近い形
態を示す。 ２．内生胞子：形成しない。 ３．運動性：認められない。 ４．鞭毛：認められない。 ５．グラム染色：陽性。 ６．コロニー：隆起している。 ７．色素：培養後期に黄色色素を産生する。 II．生理的性質 １．生育条件：20〜37℃の温度範囲で生育でき、42℃で
は生育しない。また、嫌気条件下でも生育する。 ２．硫化水素生成：陰性。 ３．インドール生成：陰性。 ４．カタラーゼ：陽性。 ５．オキシダーゼ：陰性。 ６．硝酸塩還元性：陽性であるがＮ 2 までは還元しな
い。 ７．ゼラチン液化：陽性。 ８．ウレアーゼ：陰性。 ９．アルギニンジヒドロラーゼ：陰性。 10．リジンデカルボキシラーゼ：陰性。 11．オルニチンデカルボキシラーゼ：陰性。 12．トリプトファンデアミナーゼ：陰性。 13．β−ガラクトシダーゼ：陽性。 14．糖類の資化性：グルコース、アラビノース、マンノ
ース、Ｎ−アセチル−Ｄ−グルコサミン、マルトースを
資化し、マンニトールを資化しない。 1. A possess the ability to produce significant amounts of yeast cell wall lytic enzyme and Oerusukofi <br/> A having bacteriological properties described below (Oerskovia sp.) E24 (Agency of Industrial Science Institute bacteria (Deposit No. 13692). I. Morphology and cultural properties (meat agar medium 30 ° C) 1. Morphology: At the early stage of culture, it is a long-rod bacterium and has a partially branched form
Can be seen. In the latter half of the culture, the shape is similar to monococcus to cocci
State. 2. Endospores: Do not form. 3. Motility: Not recognized. 4. Flagella: Not recognized. 5. Gram stain: positive. 6. Colony: Raised. 7. Pigment: A yellow pigment is produced in the latter stage of culture. II. Physiological properties 1. Growth conditions: Can grow in the temperature range of 20-37 ℃, at 42 ℃
Does not grow. It also grows under anaerobic conditions. 2. Hydrogen sulfide production: Negative. 3. Indole formation: negative. 4. Catalase: positive. 5. Oxidase: negative. 6. Nitrate reducibility: Positive but not reduced to N 2.
Yes. 7. Gelatin liquefaction: Positive. 8. Urease: Negative. 9. Arginine dihydrolase: negative. Ten. Lysine decarboxylase: negative. 11. Ornithine decarboxylase: negative. 12. Tryptophan deaminase: negative. 13. β-galactosidase: positive. 14. Utilization of sugars: glucose, arabinose, manno
Glucose, N-acetyl-D-glucosamine, maltose
Assimilate, not mannitol.
酵母細胞壁溶解酵素を使用することを特徴とする酵母細
胞壁の溶解方法。2. A method for lysing a yeast cell wall, which comprises using the bacterium according to claim 1 or a yeast cell wall lysing enzyme produced by the bacterium.
酵母細胞壁溶解酵素を使用することを特徴とする酵母エ
キスの製造方法。3. A method for producing a yeast extract, which comprises using the bacterium according to claim 1 or a yeast cell wall lysing enzyme produced by the bacterium.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21108193A JP3433300B2 (en) | 1993-08-04 | 1993-08-04 | Highly producing yeast cell wall lytic enzyme and method for lysing yeast cell wall using the same |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP21108193A JP3433300B2 (en) | 1993-08-04 | 1993-08-04 | Highly producing yeast cell wall lytic enzyme and method for lysing yeast cell wall using the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH0739371A JPH0739371A (en) | 1995-02-10 |
| JP3433300B2 true JP3433300B2 (en) | 2003-08-04 |
Family
ID=16600101
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP21108193A Expired - Fee Related JP3433300B2 (en) | 1993-08-04 | 1993-08-04 | Highly producing yeast cell wall lytic enzyme and method for lysing yeast cell wall using the same |
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1993
- 1993-08-04 JP JP21108193A patent/JP3433300B2/en not_active Expired - Fee Related
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| JPH0739371A (en) | 1995-02-10 |
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