JP2003153697A - Method for producing human serum albumin - Google Patents
Method for producing human serum albuminInfo
- Publication number
- JP2003153697A JP2003153697A JP2002346072A JP2002346072A JP2003153697A JP 2003153697 A JP2003153697 A JP 2003153697A JP 2002346072 A JP2002346072 A JP 2002346072A JP 2002346072 A JP2002346072 A JP 2002346072A JP 2003153697 A JP2003153697 A JP 2003153697A
- Authority
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- Japan
- Prior art keywords
- yeast
- serum albumin
- human serum
- waste liquid
- producing
- 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.)
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【０００１】[0001]
【発明の属する技術分野】本発明は、酵母菌体を含む廃
液（以下「酵母廃液」という。）を得る一方で培養物、
特に培養物から酵母菌体を分離した後の液からヒト血清
アルブミンを得ることを特徴とするヒト血清アルブミン
の製造方法に関する。TECHNICAL FIELD The present invention relates to a culture broth while obtaining a waste liquid containing yeast cells (hereinafter referred to as “yeast waste liquid”).
In particular, it relates to a method for producing human serum albumin, which comprises obtaining human serum albumin from a liquid after separating yeast cells from a culture.
【０００２】[0002]
【従来の技術】酵母菌体は、ビール、日本酒、焼酎、ウ
イスキー、ワイン、工業用エタノールなどの醸造の際、
または酵母を宿主とした組換えタンパク生産の際、副産
物として得られる。酵母菌体の一部は、医薬品、食品、
飼料などの原料として利用されるがその量は限られてお
り、余剰の酵母菌体は酵母廃液として廃棄処分されてい
る。廃棄処分の方法としては、海洋などへの投棄や焼却
処理などが行われている。しかし、環境汚染、処理コス
トの高騰などの問題から、より経済的かつ効率的な処理
方法の探索が待たれている。2. Description of the Related Art Yeast cells are used for brewing beer, sake, shochu, whiskey, wine, industrial ethanol, etc.
Alternatively, it is obtained as a by-product during recombinant protein production using yeast as a host. Some yeast cells are used in medicine, food,
It is used as a raw material for feed, etc., but its amount is limited, and surplus yeast cells are discarded as yeast waste liquid. Disposal methods include dumping into the ocean or incineration. However, due to problems such as environmental pollution and soaring treatment costs, the search for more economical and efficient treatment methods is awaited.
【０００３】[0003]
【発明が解決しようとする課題】本発明は、かかる背景
の下に、新たな処理方法を探索することによって、経済
的かつ効率的に酵母廃液の処理を行い得る処理方法を開
発し、その一方で培養物、特に培養物から酵母菌体を分
離した後の液からヒト血清アルブミンを得ることを特徴
とするヒト血清アルブミンの製造方法の提供を目的とす
る。Under the above circumstances, the present invention has developed a treatment method capable of treating yeast waste liquid economically and efficiently by searching for a new treatment method. It is an object of the present invention to provide a method for producing human serum albumin, which comprises obtaining human serum albumin from a culture, particularly a liquid after separating yeast cells from the culture.
【０００４】[0004]
【課題を解決するための手段】本発明者らは、上記課題
を解決すべく鋭意検討を重ねた結果、固定床型リアクタ
ーを用いて、酵母廃液を嫌気条件下で消化処理すること
により、経済的かつ効率的に酵母廃液の消化処理を行え
ることを見出し、本発明を完成するに到った。Means for Solving the Problems As a result of intensive studies to solve the above problems, the present inventors have found that by using a fixed bed reactor to digest yeast waste liquid under anaerobic conditions, The inventors have found that the yeast waste liquid can be digested efficiently and efficiently, and have completed the present invention.
【０００５】すなわち、本発明のヒト血清アルブミンの
製造方法は、固定床型リアクターを用いて、酵母廃液を
嫌気条件下で消化処理する、言い換えると、液化処理お
よびガス処理を含む二相式メタン発酵法により処理する
ことを特徴とする。より具体的には本発明は、ヒト血清
アルブミンを発現する組換え酵母を培地中で培養して得
られる培養物から酵母菌体を分離し、特に酵母菌体と培
養濾液とに分離した後、酵母菌体から酵母廃液を得、固
定床型リアクターを用いて、該酵母廃液を嫌気条件下で
消化処理する一方、酵母菌体を分離した後の液、例えば
酵母菌体と培養濾液とに分離した後に得られる培養濾液
からヒト血清アルブミンを得ることを特徴とするヒト血
清アルブミンの製造方法を提供する。That is, in the method for producing human serum albumin of the present invention, a fixed-bed reactor is used to digest yeast waste liquid under anaerobic conditions, in other words, two-phase methane fermentation including liquefaction and gas treatment. It is characterized by processing by the method. More specifically, the present invention separates yeast cells from a culture obtained by culturing recombinant yeast expressing human serum albumin in a medium, particularly after separating yeast cells and culture filtrate, Obtaining yeast waste liquid from yeast cells, using a fixed bed reactor, digesting the yeast waste liquid under anaerobic conditions, while separating the liquid after yeast cells are separated, for example, yeast cells and culture filtrate There is provided a method for producing human serum albumin, which comprises obtaining human serum albumin from the culture filtrate obtained after the above.
【０００６】本明細書において用いられる略語の定義は
以下の通りである。
ＴＶＳ（Total Volatile Solid） ：総揮発性物質
ＳＳ（Suspended Solid ） ：浮遊物質
ＶＳＳ（Volatile Suspended Solid）：揮発性浮遊物質
ＭＬＶＳＳ（Mixed Liquor Volatile Suspended Solid
）：混合液揮発性浮遊物質The definitions of the abbreviations used in this specification are as follows. TVS (Total Volatile Solid): Total volatile matter SS (Suspended Solid): Suspended matter VSS (Volatile Suspended Solid): Volatile suspended matter MLVSS (Mixed Liquor Volatile Suspended Solid)
): Mixed liquid volatile floating substances
【０００７】本発明において処理され得る酵母菌体の種
類は、特に限定されず、既に公知文献記載のものの他、
今後開発される組換え体などにも適宜利用することがで
きる。具体的には、サッカロマイセス属、ピキア属、シ
ゾサッカロマイセス属、クルイベロマイセス属、カンジ
ダ属、ハンゼヌラ属などが挙げられる。特に、本発明に
おいては、Ｇ４１８感受性株であるサッカロマイセス・
セレビシエ（Saccharomyces cerevisiae) ＡＨ２２株
（a. his 4, leu 2, can 1) 、ピキア・パストリス(Pic
hia pastoris) ＧＴＳ１１５株（his 4)などを宿主とし
た組換え体が好適に処理される。The type of yeast cells that can be treated in the present invention is not particularly limited, and in addition to those already known in the literature,
It can be appropriately used for recombinants to be developed in the future. Specific examples include Saccharomyces, Pichia, Schizosaccharomyces, Kluyveromyces, Candida and Hansenula. In particular, in the present invention, Saccharomyces.
Saccharomyces cerevisiae AH22 strain (a. His 4, leu 2, can 1), Pichia pastoris (Pic
Recombinant using, for example, hia pastoris GTS115 strain (his 4) as a host is preferably treated.
【０００８】これらの酵母菌体を含む酵母廃液は、生菌
の状態か、または既に公知の方法もしくはそれに準じた
方法で殺菌処理された後に、嫌気性消化により処理され
る。The yeast waste solution containing these yeast cells is sterilized by a known method or a method already known or a method similar thereto, and then processed by anaerobic digestion.
【０００９】本発明においては、固定床型リアクターを
用いて嫌気性消化を行うものである。この方法は、別
名、固定床法（ＵＡＦＰ：Upflow Anaerobic Filter Pr
ocess）ともいう。固定床法は、固定化担体に嫌気性微
生物を付着させた固定床型リアクターにより消化処理を
行うというものである。この固定化担体としては、セラ
ミック、ポリアクリルアミド、砕石等が例示される。本
発明においては、少なくとも液化処理装置として固定床
型リアクターを利用する。In the present invention, anaerobic digestion is carried out using a fixed bed reactor. This method is also known as the fixed bed method (UAFP: Upflow Anaerobic Filter Pr).
Also called ocess). The fixed bed method is to perform digestion treatment by a fixed bed reactor in which anaerobic microorganisms are attached to an immobilized carrier. Examples of the immobilization carrier include ceramics, polyacrylamide, and crushed stone. In the present invention, a fixed bed reactor is used as at least a liquefaction treatment device.
【００１０】消化処理に適用される酵母廃液のＴＶＳ濃
度は、１０〜１００ｇ−ＴＶＳ／Ｌが望ましい。酵母廃
液のＴＶＳ濃度が１０ｇ−ＴＶＳ／Ｌ未満では、水理学
的滞留時間が短くなり、ＶＳＳ消化率が低くなるなど効
率的な嫌気性消化、特に液化処理が困難となる。一方、
１００ｇ−ＴＶＳ／Ｌを越えると、プロピオン酸などの
有機酸の生成量が多くなり、以後のガス化処理に悪影響
を及ぼす。The TVS concentration of the yeast waste liquid applied to the digestion treatment is preferably 10 to 100 g-TVS / L. When the TVS concentration of the yeast waste liquid is less than 10 g-TVS / L, the hydraulic retention time becomes short and the VSS digestibility becomes low, so that efficient anaerobic digestion, particularly liquefaction treatment becomes difficult. on the other hand,
When it exceeds 100 g-TVS / L, the amount of organic acids such as propionic acid produced increases, which adversely affects the subsequent gasification treatment.
【００１１】液化処理において作用する微生物は、酸生
成菌と水素生成酢酸生成菌とに大きく分けられる。酸生
成菌は、高分子有機物から酢酸、プロピオン酸、酪酸な
どの低級脂肪酸を生成する一群の微生物である。また、
水素生成酢酸生成菌は、水素資化性のメタン生成菌や硫
酸還元菌と共生して低級脂肪酸から酢酸と水素とを生成
する。当該微生物としては、下記の表１に記載のものが
例示される。Microorganisms that act in the liquefaction process are roughly classified into acid-producing bacteria and hydrogen-producing acetic acid-producing bacteria. Acid-producing bacteria are a group of microorganisms that produce lower fatty acids such as acetic acid, propionic acid, and butyric acid from macromolecular organic substances. Also,
The hydrogen-producing acetic acid-producing bacterium coexists with hydrogen-utilizing methanogenic bacteria and sulfate-reducing bacteria to produce acetic acid and hydrogen from lower fatty acids. Examples of the microorganism include those listed in Table 1 below.
【００１２】[0012]
【表１】 [Table 1]
【００１３】ガス化処理において作用する嫌気性菌は、
Methanobacteriales, Methanococcales, Methanomicrob
iales の３目に分けられ、さらに下記の表２に示される
４科８属１４種に分類される。Anaerobic bacteria that act in gasification treatment are
Methanobacteriales, Methanococcales, Methanomicrob
It is divided into 3 subgroups of iales, and is further classified into 14 species of 4 families and 8 genera shown in Table 2 below.
【００１４】[0014]
【表２】 [Table 2]
【００１５】これらの嫌気性菌のうち、Methanosarcina
属とMethanothrix属とは酢酸を基質にできるメタン生成
菌であり、この２属がメタン生成過程の主要な役割を果
している。Among these anaerobic bacteria, Methanosarcina
Genus Methanothrix is a methanogenic bacterium that can use acetic acid as a substrate, and these two genera play a major role in the methanogenic process.
【００１６】嫌気性消化処理は、種々雑多の当該微生物
を含む（微生物群からなる）汚泥または嫌気性消化シー
ド〔種汚泥または嫌気性消化汚泥ともいう。下水処理な
どにおいて得られた嫌気性汚泥（嫌気性菌群）を馴養し
たもの。〕を通常の方法により馴養する。すなわち、こ
の馴養は嫌気性消化処理を行い易い状態に馴らしていく
ことを意味する。具体的には、嫌気性消化シードを例え
ば、ＭＬＶＳＳとして１〜５０ｇ／Ｌの濃度に対して、
混合汚泥、合成廃水などを用いて、有機物容積負荷、ｐ
Ｈ、温度条件などの条件を当該液化処理の条件に合わせ
た条件下で適当な期間馴養する。次いで、酵母廃液の嫌
気性消化処理を開始する。The anaerobic digestion treatment is a sludge containing various miscellaneous microorganisms (comprising a group of microorganisms) or an anaerobic digestion seed [also referred to as seed sludge or anaerobic digestion sludge. An anaerobic sludge (anaerobic bacteria group) obtained by sewage treatment, etc. ] Is acclimated by the usual method. That is, this acclimatization means acclimatization to a state where anaerobic digestion treatment is easily performed. Specifically, for anaerobic digestion seeds, for example, MLVSS as a concentration of 1 to 50 g / L,
Using mixed sludge, synthetic wastewater, etc., organic matter volume loading, p
The conditions such as H and temperature conditions are acclimated for an appropriate period under conditions that match the conditions of the liquefaction process. Then, the anaerobic digestion treatment of the yeast waste liquid is started.
【００１７】また、複数のリアクターを用いる場合は、
その各々のリアクターで行う嫌気性消化処理（例えば、
液化処理、ガス化処理）を行い易い状態に馴養すればよ
い。When using a plurality of reactors,
Anaerobic digestion process (eg,
The liquefaction process and the gasification process) should be acclimated to a state where it is easy to perform.
【００１８】酵母廃液は希釈せずに、あるいは水または
何らかの水溶液で希釈した上で、当該処理を行う。当該
処理は公知の温度条件下で行うことができる。一般的に
は３０〜７０℃程度が例示される。特に本発明において
は、中温発酵（３０〜４０℃）および高温発酵（５０〜
６０℃）のいずれでも実施可能である。The yeast waste liquid is not diluted, or diluted with water or some aqueous solution, and then the treatment is carried out. The treatment can be performed under known temperature conditions. Generally, about 30 to 70 ° C. is exemplified. Particularly in the present invention, medium temperature fermentation (30 to 40 ° C.) and high temperature fermentation (50 to
Any of 60 ° C.) is possible.
【００１９】当該処理は連続して行うことができる。有
機物容積負荷は、０．５〜１０ｇ−ＴＶＳ／Ｌ／日程
度、好ましくは１〜８ｇ−ＴＶＳ／Ｌ／日程度とする。
ｐＨは６〜８程度となるように調整すればよく、酸また
はアルカリを適宜添加して制御する。The treatment can be performed continuously. The organic substance volume load is about 0.5 to 10 g-TVS / L / day, preferably about 1 to 8 g-TVS / L / day.
The pH may be adjusted so as to be about 6 to 8 and is controlled by appropriately adding an acid or alkali.
【００２０】当該処理においては、必要に応じてニッケ
ルおよび／またはコバルトの塩を酵母廃液に添加するこ
とが、消化反応を促進させる点で好ましい。具体的に
は、塩化第一ニッケル六水塩、塩化第一コバルト六水塩
などが例示される。ニッケルおよび／またはコバルトの
塩は、酵母廃液のＴＶＳ濃度に応じて適宜添加される。
具体的には、終濃度として各々０．５〜１００mg／Ｌ程
度が例示される。In the treatment, it is preferable to add a salt of nickel and / or cobalt to the yeast waste solution, if necessary, from the viewpoint of promoting the digestion reaction. Specific examples thereof include nickel (I) chloride hexahydrate and cobalt (I) chloride hexahydrate. The nickel and / or cobalt salt is appropriately added depending on the TVS concentration of the yeast waste liquid.
Specifically, the final concentration is, for example, about 0.5 to 100 mg / L.
【００２１】当該処理条件下で、酵母廃液は連続的に処
理され、例えば液化処理工程におけるＶＳＳの消化率を
５０％以上に維持することが可能となる。ガス化処理も
安定して行うことができ、生成されるガスは５０％以上
のメタンを含有し、燃料などとして利用することができ
る。Under the processing conditions, the yeast waste liquid is continuously processed, and for example, the digestibility of VSS in the liquefaction processing step can be maintained at 50% or more. The gasification process can be performed stably, and the produced gas contains 50% or more of methane and can be used as a fuel or the like.
【００２２】複数のリアクターを連続して用いる場合
は、得られた消化処理液を希釈せずに、あるいは水また
は何らかの水溶液で希釈して、次のリアクター（処理工
程）に投入される。When a plurality of reactors are continuously used, the obtained digestion treatment liquid is not diluted, or diluted with water or some kind of aqueous solution, and then fed to the next reactor (treatment step).
【００２３】なお、ガス化処理を液化処理とは別のリア
クターで行う場合は、良好なガス発生量（４００ml-gas
／ｇ−ＴＶＳ以上）を達成するために、液化処理液を揮
発性脂肪酸（ＶＦＡ）濃度１０ｇ／Ｌ以下となるように
調整して用いることが好ましい。When the gasification process is carried out in a reactor different from the liquefaction process, a good gas generation amount (400 ml-gas
/ G-TVS or more), it is preferable that the liquefaction treatment liquid is adjusted to a volatile fatty acid (VFA) concentration of 10 g / L or less before use.
【００２４】[0024]
【実施例】以下、本発明をより詳細に説明するために実
施例を挙げるが、本発明はこれら実施例に何ら限定され
るものではない。EXAMPLES Examples will be given below to explain the present invention in more detail, but the present invention is not limited to these examples.
【００２５】実施例１
酵母廃液の調製
ピキア・パストリス(Pichia pastoris) ＧＴＳ１１５株
に、ヒト血清アルブミン発現用プラスミドｐＭＭ０４２
を導入し、形質転換体ＵＨＧ４２−３株を得た（特開平
４−２９９８４号公報参照）。Example 1 Preparation of yeast waste liquor Pichia pastoris GTS115 strain was added to plasmid pMM042 for expression of human serum albumin.
Was introduced to obtain a transformant UHG42-3 strain (see JP-A-4-29984).
【００２６】この形質転換体ＵＨＧ４２−３株をジャー
ファーメンターを用いて通気攪拌培養し、培養終了後に
圧搾機にて、酵母菌体と濾液とに分離した。圧搾により
得られた酵母菌体を湿重で２５０ｇ採取し、これを蒸留
水にて懸濁して１Ｌの懸濁液とした。この懸濁液を１２
１℃にて２０分間蒸気加圧滅菌して、酵母廃液１Ｌを得
た。この酵母廃液にＮｉＣｌ2・６Ｈ2ＯおよびＣｏＣｌ
2・６Ｈ2Ｏをそれぞれ終濃度で１８．１および５．１mg
／Ｌとなるように添加した。The transformant UHG42-3 strain was cultivated with aeration and stirring using a jar fermenter, and after completion of the culturing, yeast cells and the filtrate were separated by a pressing machine. 250 g of the yeast cells obtained by pressing were collected under wet weight and suspended in distilled water to give a 1 L suspension. This suspension 12
Steam sterilization was performed at 1 ° C. for 20 minutes to obtain 1 L of yeast waste liquid. NiCl 2 .6H 2 O and CoCl were added to this yeast waste solution.
18.1 and 5.1mg 2 · 6H 2 O at each final concentration
/ L.
【００２７】この酵母廃液の浮遊物質（ＳＳ）濃度は、
約65,000mg／Ｌ、揮発性浮遊物質（ＶＳＳ）濃度は約6
0,000mg／Ｌ、総揮発性物質（ＴＶＳ）濃度は約65,000m
g／Ｌであった。The suspended solids (SS) concentration of this yeast waste solution is
Approximately 65,000 mg / L, volatile suspended solids (VSS) concentration is approximately 6
0,000mg / L, total volatile substance (TVS) concentration is about 65,000m
It was g / L.
【００２８】 消化シードの馴養
高温消化シード（嫌気性植種汚泥）を有機物容積負荷３
ｇ／Ｌ／日の条件下で、下水混合汚泥を用いてdraw-and
-fill 方式により馴養した。Acclimation of Digestion Seeds High-temperature digestion seeds (anaerobic seed sludge) were added to the organic matter volume load 3
draw-and using sewage mixed sludge under the condition of g / L / day
-According to the fill method.
【００２９】 嫌気性消化処理
０．７８Ｌ容の固定床型リアクターに、馴養した高温消
化シードをＭＬＶＳＳで１０ｇ／Ｌとなるように投入し
た。なお、固定床型リアクターに充填した担体は、リア
ポア（粘土を焼いて粉砕したもの）と粉砕した高密度ポ
リエチレンとを４５：５５に混合し、１００〜２３０℃
でハニカム構造に焼成したものである。従って、この担
体は、親水的な性質を（リアポア）と疎水的な性質（高
密度ポリエチレン）とを併せ持っている。この担体は非
常にポーラスであり、細孔分布は１〜２mmの細孔が２０
％、０．１〜０．３５mmの細孔が６０％、０．１μｍ〜
０．１mmの細孔が２０％である。Anaerobic digestion treatment A fixed-bed reactor having a volume of 0.78 L was charged with the acclimated hot digestion seeds so as to have an MLVSS concentration of 10 g / L. The carrier packed in the fixed bed reactor was a mixture of rear pores (clay baked and crushed) and crushed high density polyethylene at 45:55, and the mixture was heated at 100 to 230 ° C.
The honeycomb structure is fired. Therefore, this carrier has both a hydrophilic property (rear pore) and a hydrophobic property (high density polyethylene). This carrier is very porous and has a pore size distribution of 1 to 2 mm.
%, 60% of 0.1-0.35 mm pores, 0.1 μm-
20% of 0.1 mm pores.
【００３０】槽内液は循環ポンプで循環した。また、酵
母廃液は３倍希釈したものを供給ポンプにより連続的に
供給した。処理温度は５３℃、有機物容積負荷は２ｇ／
Ｌ／日とし、嫌気性消化処理を５０日間行った。The liquid in the tank was circulated by a circulation pump. The yeast waste liquid was diluted three-fold and continuously supplied by a supply pump. Treatment temperature is 53 ° C, organic volume loading is 2g /
L / day, and anaerobic digestion treatment was performed for 50 days.
【００３１】実験例１
実施例１において、有機物容積負荷を２〜８ｇ−ＴＶＳ
／Ｌ／日の条件下で、嫌気性消化処理を行った場合の結
果を表３に示す。Experimental Example 1 In Example 1, the organic matter volume load was 2 to 8 g-TVS.
Table 3 shows the results when the anaerobic digestion treatment was performed under the condition of / L / day.
【００３２】[0032]
【表３】 [Table 3]
【００３３】実施例２
中温消化シードを用い、処理温度を３７℃とする以外は
実施例１に準じて酵母廃液の嫌気性消化処理を行った。
その結果、実施例１と同様の良好な結果が得られた。Example 2 An anaerobic digestion treatment of yeast waste liquid was carried out in the same manner as in Example 1 except that a medium temperature digestion seed was used and the treatment temperature was 37 ° C.
As a result, the same good result as in Example 1 was obtained.
【００３４】実施例３
酵母廃液の加熱滅菌を６８℃で３０分間行い、酵母廃液
のｐＨを６．５に調整する以外は実施例１に準じて酵母
廃液の嫌気性消化処理を行った。その結果、実施例１と
同様の良好な結果が得られた。Example 3 The yeast waste liquid was subjected to anaerobic digestion treatment in the same manner as in Example 1 except that the yeast waste liquid was heat-sterilized at 68 ° C. for 30 minutes and the pH of the yeast waste liquid was adjusted to 6.5. As a result, the same good result as in Example 1 was obtained.
【００３５】実験例２
実施例３において、有機物容積負荷を２〜４ｇ−ＴＶＳ
／Ｌ／日の条件下で、嫌気性消化処理を行った場合のＶ
ＳＳ消化率を表４に示す。Experimental Example 2 In Example 3, the organic matter volume load was 2 to 4 g-TVS.
V under anaerobic digestion treatment under the condition of / L / day
Table 4 shows the SS digestibility.
【００３６】[0036]
【表４】 [Table 4]
【００３７】[0037]
【発明の効果】本発明のヒト血清アルブミンの製造方法
によれば、ヒト血清アルブミンを得る一方で酵母廃液を
固定床型リアクターを用いて、嫌気条件下で消化処理を
行うことができる。かかる処理により酵母菌体を含む酵
母廃液が高効率的にメタンガスに転換され、酵母廃液の
経済的な廃棄処理が可能となる。According to the method for producing human serum albumin of the present invention, while the human serum albumin is obtained, the yeast waste solution can be digested under anaerobic conditions by using the fixed bed reactor. By such a treatment, the yeast waste liquid containing the yeast cells is highly efficiently converted to methane gas, and the yeast waste liquid can be economically disposed of.
【００３８】特に、本発明の方法によれば、酵母廃液の
廃棄処理において、従来に比べて、有機物容積負荷をよ
り高く設定することができる。また、前処理としての加
熱処理は６０〜７０℃程度であっても、嫌気性消化反応
を充分に行うことができる。In particular, according to the method of the present invention, it is possible to set the organic matter volume load higher in the waste treatment of the yeast waste liquid than in the conventional case. Moreover, even if the heat treatment as the pretreatment is about 60 to 70 ° C., the anaerobic digestion reaction can be sufficiently performed.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 桑江 忍 大阪府枚方市招提大谷２丁目25番１号 株 式会社ミドリ十字中央研究所内 (72)発明者 應田 豊雄 大阪府枚方市招提大谷２丁目25番１号 株 式会社ミドリ十字中央研究所内 (72)発明者 大村 孝男 大阪府枚方市招提大谷２丁目25番１号 株 式会社ミドリ十字中央研究所内 Ｆターム(参考） 4B064 AG01 CA06 CA19 CC24 DA01 ─────────────────────────────────────────────────── ─── Continued front page (72) Inventor Shinobu Kuwae 2-25-1 Otani Invitation, Hirakata City, Osaka Prefecture Ceremony company Midori Cross Central Research Institute (72) Inventor Toyo Ohta 2-25-1 Otani Invitation, Hirakata City, Osaka Prefecture Ceremony company Midori Cross Central Research Institute (72) Inventor Takao Omura 2-25-1 Otani Invitation, Hirakata City, Osaka Prefecture Ceremony company Midori Cross Central Research Institute F-term (reference) 4B064 AG01 CA06 CA19 CC24 DA01
Claims (4)
母を培地中で培養して得られる培養物から酵母菌体を分
離した後、酵母菌体から酵母廃液を得、固定床型リアク
ターを用いて、該酵母廃液を嫌気条件下で消化処理する
一方、酵母菌体を分離した後の液からヒト血清アルブミ
ンを得ることを特徴とするヒト血清アルブミンの製造方
法。1. A method for culturing recombinant yeast expressing human serum albumin in a medium, separating yeast cells from a culture product, obtaining yeast waste liquid from the yeast cells, and using a fixed bed reactor. A method for producing human serum albumin, which comprises digesting the yeast waste liquid under anaerobic conditions and obtaining human serum albumin from the liquid after separating yeast cells.
て、当該製造過程において、ヒト血清アルブミンを発現
する組換え酵母を培地中で培養して得られる培養物から
酵母菌体を分離した後、酵母菌体から酵母廃液を得、固
定床型リアクターを用いて、該酵母廃液を嫌気条件下で
消化処理する工程を有することを特徴とするヒト血清ア
ルブミンの製造方法。2. A method for producing human serum albumin, which comprises separating a yeast cell from a culture obtained by culturing a recombinant yeast expressing human serum albumin in a medium in the production process, and then producing the yeast. A method for producing human serum albumin, which comprises a step of obtaining a yeast waste liquid from cells and digesting the yeast waste liquid under anaerobic conditions using a fixed bed reactor.
母を培地中で培養して得られる培養物を酵母菌体と培養
濾液とに分離した後、酵母菌体を液体中に懸濁して酵母
廃液を得、固定床型リアクターを用いて、該酵母廃液を
嫌気条件下で消化処理する一方、培養濾液からヒト血清
アルブミンを得ることを特徴とするヒト血清アルブミン
の製造方法。3. A yeast waste liquid obtained by culturing a recombinant yeast expressing human serum albumin in a medium, separating a culture product into yeast cells and a culture filtrate, and suspending the yeast cells in a liquid. And a method of producing human serum albumin from a culture filtrate while digesting the yeast waste solution under anaerobic conditions using a fixed bed reactor.
て、当該製造過程において、ヒト血清アルブミンを発現
する組換え酵母を培地中で培養して得られる培養物を酵
母菌体と培養濾液とに分離した後、酵母菌体を液体中に
懸濁して酵母廃液を得、固定床型リアクターを用いて、
該酵母廃液を嫌気条件下で消化処理する工程を有するこ
とを特徴とするヒト血清アルブミンの製造方法。4. A method for producing human serum albumin, wherein in the production process, a culture obtained by culturing recombinant yeast expressing human serum albumin in a medium is separated into yeast cells and a culture filtrate. After that, the yeast cells are suspended in a liquid to obtain a yeast waste liquid, and using a fixed bed reactor,
A method for producing human serum albumin, which comprises a step of digesting the yeast waste liquid under anaerobic conditions.
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|---|---|---|---|
| JP31038995A Division JP3743039B2 (en) | 1995-11-29 | 1995-11-29 | Yeast waste liquid treatment method |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009017847A (en) * | 2007-07-13 | 2009-01-29 | Nipro Corp | Method for producing cell culture medium, and cell culture medium |
| CN111662944A (en) * | 2019-03-05 | 2020-09-15 | 上海医药工业研究院 | Preparation method and purification method of human serum albumin |
-
2002
- 2002-11-28 JP JP2002346072A patent/JP2003153697A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009017847A (en) * | 2007-07-13 | 2009-01-29 | Nipro Corp | Method for producing cell culture medium, and cell culture medium |
| CN111662944A (en) * | 2019-03-05 | 2020-09-15 | 上海医药工业研究院 | Preparation method and purification method of human serum albumin |
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