JP2696530C

JP2696530C -

Info

Publication number: JP2696530C
Authority: JP
Original assignee: 東和化成工業株式会社
Publication date: 1999-08-30

Description

【発明の詳細な説明】（産業上の利用分野）本発明はオリゴ糖の少ないマルトースの製造方法及びその還元物の製造方法に
関するものである。（従来の技術）マルトース、即ち４−〔α−Ｄ−グルコピラノシル〕−Ｄ−グルコースは古くから麦芽水飴の主成分として知られ、良質の風味
を有するために広く食品に使用されてきた。一方、その還元物であるマルチトー
ル、即ち４−〔α−Ｄ−グルコピラノシル〕−Ｄ−グルチトールも微生物により
醗酵されにくいことや、砂糖に近い甘味を呈することなどの利点を有することか
ら食品、化粧品、薬品などの分野で広範囲の用途に使用されている。しかし、最近食品用のマルトース又はマルチトールとして粉末品の需要が増大
しており、更に医薬品用途のマルトースやマルチトールは高純度の品が要望され
ており、一方では粉末品のなかでも吸湿性の低いものが望まれている。また、マ
ルトース又はマルチトールの粉末品はマルトース又はマルチトールの純度が高い
ものほど粉末化が容易であるという技術的事情もあった。以上の背景からマルトース又はマルチトールの粉末品を作る場合には、その純
度を高めることによって粉末化させようとした試みが主流になり、多くの方法が
紹介されている。しかしながら、それらの方法はマルトースやマルチトールの純度を高めるため
に糖化工程の管理が極めて困難であったり、特殊な酵素を使用しているために経
済的に不利であったり、クロマト分画などの困難で手間のかかる工程が含まれて
いたりなどの工業的に実施する上で不都合な点があった。その後、これらの不都
合はマルトース又はマルチトールの純度を極めて高くして粉末化を容易にしよう
としたために発生したものであることに着目し、その改善方法が検討された。そ
の結果、たとえ単糖又は単糖の還元物が少々増加したとしても、オリゴ糖又はオ
リゴ糖アルコールを少なくしようとする試みがなされた。その改善方法としては
、特公昭57-3356 号公報や特公昭56-28153号公報、特公昭56-28154号公報
、更に特願昭63-101355 号や特願昭63-101356 号に記載されているような方
法などがあり一応の成果を見ている。上記の改善方法、、の要点は、糖化
の際にマルトトリオース分解活性／マルトース分解活性≧2.5である酵素等を作
用させるというものであり、改善方法及びの要点は、汎用性の高い酵素を特殊な組み合わせで
使用して特定の組成の粉末化の容易な糖液を調製するというものであった。（発明が解決しようとする課題）しかしながら従来の方法には依然として課題が残されており、工業的に粉末化
の容易なマルトース又はマルチトールを製造する方法として充分に有利な方法と
はいえなかった。例えば、前記、、の方法は、糖化の際に使用する酵素として特殊なもの
を使用しているために、酵素の入手が困難であるという不都合があった。更に糖
化の際にマルトースも比較的に多く加水分解されるためにグルコース含量が増加
してマルトース収率が高まらないので、開示されている各種条件の中でもデキス
トロース当量（ＤＥ）１前後で液化を止めて糖化し、高純度マルトースを製造す
る必要があった。つまりＤＥ１前後で液化を止めるという極めて困難な方法であ
った。また、前記、の方法は工程が比較的長く、純度の高いマルトース又はマルチトールを得ようとしたときはクロマト分離等の
条件が比較的正確なものを要求されるなどの課題を有していた。そのために、従来の方法は粉末化の容易な、且つオリゴ糖の少ないマルトース
及びその還元物を工業的に有利に製造する方法として十分なものではなかった。以上のような状況から工業的にオリゴ糖の少ないマルトース及びその還元物を
製造するための、より容易でしかも有利な方法の開発が強く望まれていた。（課題を解決するための手段）上記諸々の課題を解決するために、本発明者等は鋭意研究を重ねた結果、バチ
ルス・ステアロサーモフィルス(Bacillus stearothermophilus)の遺伝子のマル
トゲニック−α−アミラーゼがコードされた部分をプラスミドにはめ込み、バチ
ルス・ズブティリス(Bacillus subtilis)に組込んで生産されたマルトゲニック
−α−アミラーゼ（以下単にマルトゲニックアミラーゼということがある。）を澱粉液化物の糖化の際に特定の方法で使用することによって、従来の方
法よりも簡単な工程で、容易に粉末化可能なオリゴ糖の少ないマルトース及びそ
の還元物を製造することに成功し、本発明を完成するに至った。以下に本発明の内容を詳細に説明する。本発明の目的は簡単な工程で、入手しやすい酵素を使用し、経済的に有利な地
上澱粉をも利用可能にし、粉末化の容易な、オリゴ糖含有量の少ないマルトース
及びその還元物を製造する方法を提供することにある。即ち、第１番目の発明は濃度５〜４０重量％の澱粉水溶液に液化酵素を作用
させて液化し、デキストロース当量１５以下にて液化酵素を失活させる第１工程
、上記工程で得られた液化物にβ−アミラーゼ及びプルラナーゼ及び／又はイ
ソアミラーゼを作用させると同時に又は作用開始後３６時間以内にマルトゲニッ
クアミラーゼを作用させて糖化する第２工程、第２工程開始後、１〜４８時間後に液化酵素を基質固形分１ｇあたり１〜２
０単位添加して更に糖化し、糖化物のマルトース含量が固形分中７５〜９０重量
％で、且つ糖化物に含まれるオリゴ糖の含有量が次式で計算したとき７以下の数
値の範囲に糖化する第３工程、から構成される。又、第２番目の発明は濃度５〜４０重量％の澱粉水溶液に液化酵素を作用さ
せて液化し、デキストロース当量１５以下にて液化酵素を失活させる第１工程、上記工程で得られた液化物にβ−アミラーゼ及びプルラナーゼ及び／又はイ
ソアミラーゼを作用させると同時に又は作用開始後３６時間以内にマルトゲニッ
クアミラーゼを作用させて糖化する第２工程、第２工程開始後、１〜４８時間後に液化酵素を基質固形分１ｇあたり１〜２
０単位添加して更に糖化し、糖化物のマルトース含量が固形分中７５〜９０重量％で、且つ糖化物
に含まれるオリゴ糖の含有量が次式で計算したとき７以下の数値の範囲に糖化す
る第３工程、第３工程で得られた糖化物を還元する第４工程、により構成される。上記各発明の工程を経由して得られたオリゴ糖の少ないマルトース又はその還
元物は、公知の方法により、精製、濃縮、クロマト分離、結晶化（固化）、乾燥
、粉末化などの工程に供することによって容易に粉末状又は結晶状のマルトース
又はマルチトールを、更に高純度のマルトース又はマルチトールを調製すること
ができる。以下に、本発明の内容を更に詳細に説明するが、最初に、第１番目の発明の内
容を説明する。本発明には地上澱粉、地下澱粉の別を問わず使用可能であるが、特に従来は粉
末状又は高純度マルトース及びその還元物を製造するうえで不都合の多かった地上澱粉も有利に使
用可能であることが、本発明の利点の一つである。本発明を実施するうえでこの
澱粉中のアミロースやアミロペクチンの組成も特に気にする必要はなく、使用可
能な澱粉を具体的に例示すると、トウモロコシ澱粉、小麦澱粉、大麦澱粉、など
の地上澱粉の他に各種の地下澱粉があげられる。これらの澱粉を液化する際、地上澱粉を原料としたときは特に液化液の老化を
防ぐ意味で液化時の基質濃度を好ましくは１０〜３０％、pHを6.0〜6.8に調整し
て耐熱性の液化酵素例えばノボ社のターマミル（登録商標）などの液化酵素を使
用して液化し、デキストロース当量１５以下、更に好ましくは３〜１３で液化酵
素を失活させることが望ましい。次に、液化物にβ−アミラーゼ、プルラナーゼ及び／又はイソアミラーゼ及び
マルトゲニックアミラーゼをβ−アミラーゼ及びプルラナーゼ及び／又はイソア
ミラーゼと同時〜３６時間以内に作用させて糖化するが、その一般的な好ましい条件は基質濃度５〜４０重量％、pH
5.3、温度５５℃程度である。この糖化開始後１〜４８時間後に液化酵素を基質固形分１ｇあたり１〜２０単
位添加して更に糖化するが、この操作により、主に四糖以上のオリゴ糖を加水分
解してマルトース及び三糖を生成し、必要に応じてその後に行われる工程の一層
の効果発現を促す糖組成とし、ろ過性を改善することができる。この時に使用する酵素は、β−アミラーゼとしては長瀬産業（株）製のβ−ア
ミラーゼ＃１５００、フィンシュガー社製のスペザイム（SPEZYME;登録商標）BB
A 1500などがあるが、それらの中でも大豆由来のβ−アミラーゼが本発明を実施
するうえで有利な性質をそなえており、プルラナーゼとしてはノボ社のプロモザ
イムや天野製薬(株)製のプルラナーゼアマノＣＫＬ等が汎用性が高く、市販され
ていることや酵素の性質等から有利に使用できる。一方、前記の遺伝子組変えにより製造されたマルトゲニックアミラーゼとして
は、ノボ社のマルトゲナーゼ（Maltogenase；登録商標）がある。糖化のときに使用する好ましい酵素量の比率は、例えば前記遺伝子組変えによ
り製造したノボ社のマルトゲナーゼを１〜２０単位（このマルトゲナーゼの活性
はマルトゲニック・アミラーゼ・ノボ・ユニットを採用して説明する。）使用し
たときにβ−アミラーゼが１０〜３０単位、プルラナーゼが0.6〜2.0単位である
。また、この糖化工程はマルトースの純度が平衡に達するまで（通常２４〜７２
時間）を目安に行う。この第３工程で使用する液化酵素は非耐熱性でも耐熱性でも使用可能であるが
、非耐熱性の液化酵素の方が本発明を実施するうえで一層効果的である。以上の工程によってマルトース純度７５〜９０重量％で、且つ糖化物に含まれ
るオリゴ糖の含有量が次式で計算したとき７以下の数値の範囲に糖化することが
できる。この工程によって得られる糖化物は現在市販されているマルトースを主成分と
する製品群のなかでは比較的高いマルトース純度を有するものであり、その成分
組成は三糖以上のオリゴ糖含有量が少ないので、市販の類似製品に比較して粘度
が低く、結晶化した場合にはマルトースの結晶成長速度が速いので、公知の方法
で直接結晶・粉末化することが容易である。更に、晶析やクロマト分離工程に供
して純度を高めようとした場合にも従来の方法で製造した製品よりも一層有利に
高純度のマルトースを製造することができる。粉末マルトースを製造する方法としては、例えば分蜜法、噴霧造粒法、流動造
粒法、ブロック粉砕法の各種方法またはそれらの組み合わせが採用可能である。必要に応じてマルトース純度を高めるときは、イオン交換樹脂、イオン交換繊
維、ゼオライトなどの各種イオン交換体をアルカリ金属型にしてクロマト分離する方法や適切な濃度まで濃縮した後、晶析、分蜜化する方法などが
採用可能である。次に、第２番目の発明の工程を説明する。第２番目の発明は、第１番目の発明で得られた糖化物をそのまま又は必要に応
じて上記各種手段でマルトース純度を高めた後、それ自身は公知の回分式または
連続式の方法で、ニッケル系または貴金属系などの還元触媒の存在下で水素添加
してマルチトールを主成分とする糖アルコールにする。この水素添加条件はマル
トースの分解が生じない条件であれば、どのような条件でも良いが、通常は糖液
の濃度を４０〜６０重量％にして、水素圧２０kg／cm²以上、更に好ましくは５
０〜２００kg／cm²で、１００〜１５０℃の温度で行う。この工程によって得られるオリゴ糖含有量の少ないマルチトールは、現在市販
されているマルチトールを主成分とする製品群の中では比較的高いマルチトール
純度を有するものであり、その成分組成は三糖以上のオリゴ糖アルコール含有量
が少ないので、市販の類似製品に比較して粘度が低く、結晶化した場合にはマルチトールの結晶成長速度が速いので、公知の方法で直接
結晶・粉末化することが容易である。この後必要に応じてクロマト分離法や晶析分蜜化などの公知の方法によって更
にマルチトールの純度を高めることも容易に可能である。（実施例）次に実施例を掲げて本発明の内容を更に具体的に説明するが、本発明は以下の
実施例によって限定されるものではない。実施例−１第１工程（液化工程）トウモロコシ澱粉を濃度３０％、pH6.2に調整し、耐熱液化酵素〔長瀬産業(株
)製、スピターゼHS〕２０ｕ／ｇ基質固形分（以下DSと略する。）を添加して常
法にて１０５℃で液化した。加熱により液化反応をＤＥ6.5にて停止させた。第２工程次に、温度５５℃、pH5.3に調整した後、１０ｕ／ｇＤＳの長瀬産業(株)製β
−アミラーゼ＃1500 及び0.67ｕ／ｇＤＳのノボ社製プロモザイムＴＭ２００Ｌを添加して糖化反応を
進めた。第２工程開始後６時間目にノボ社製マルトゲナーゼ6.5ｕ／ｇＤＳを添加した
。第３工程更に第２工程開始後６時間目に上記スピターゼＰＮ−４を２０ｕ／ｇＤＳ添加
して更に６６時間糖化反応を継続した。第３工程終了後の糖組成を高速液体クロマトグラフィーにて測定した結果は次
の通りであった。一糖 7.0％二糖 88.2％三糖 1.0％四糖以上のオリゴ糖 3.8％実施例−２実施例−１で得た糖化物を常法に従って脱色、脱塩、濃縮して濃度５０％の精
製糖液とし、その２０kgとラネーニッケル触媒２００ｇを内容積２５ｌのオート
クレーブに仕込み、水素圧を150〜１２０kg／cm²に保ち、１２０℃にて２時間攪拌し、水素添加を行った。得られた液を触媒と分離した後、粒状活性炭のカラム
を通して高速液体クロマトグラフィーにて分析した結果は以下の通りであった。ソルビトール 7.4％マルチトール 88.1％三糖以上のオリゴ糖アルコール 4.5％得られた還元物を精製処理後、濃度７５％まで濃縮して１０℃に冷却後、種晶
３重量％を添加混合して攪拌しながら１５時間かけてマスキットを調製し、噴霧
乾燥機で送風温度８０℃にて噴霧結晶化し、結晶状の粉末マルチトールを得た。比較例−１比較のために市販のマルトース液（糖組成＝一糖1.4％、二糖90.0％、三糖7.0
％、四糖以上のオリゴ糖1.6％）を還元し、下記の組成を有する還元物を得た。ソルビトール 1.5％マルチトール 89.6％三糖以上のオリゴ糖アルコール 8.9％これに実施例−２と同様に精製・濃縮冷却後マスキット調製操作を行ったが１
５時間後噴霧乾燥可能なマスキットは得られなかった。（発明の効果）以上に述べたように、本発明を実施することにより、比較的短い工程で、容易
な工程管理で、経済的に有利に、容易に粉末化可能な組成の、オリゴ糖の少ない
マルトース及びその還元物を製造することが可能になる。Description: TECHNICAL FIELD The present invention relates to a method for producing maltose having a small amount of oligosaccharide and a method for producing a reduced product thereof. (Prior art) Maltose, that is, 4- [α-D-glucopyranosyl] -D-glucose, has long been known as a main component of malt syrup, and has been widely used in foods because of its high quality flavor. On the other hand, its reduced product, maltitol, that is, 4- [α-D-glucopyranosyl] -D-glutitol also has advantages such as being hardly fermented by microorganisms and having a sweetness similar to sugar, so that it can be used in foods and cosmetics. It is used for a wide range of applications in the fields of medicine, medicine and the like. However, recently, the demand for powdered maltose or maltitol for food is increasing, and maltose or maltitol for pharmaceutical use is required to be of high purity. A lower one is desired. In addition, there is also a technical situation that the powdery maltose or maltitol has a higher purity of maltose or maltitol, so that powdering is easier. In view of the above background, in the case of producing a powdery product of maltose or maltitol, attempts to increase the purity of the maltose or maltitol have become mainstream, and many methods have been introduced. However, these methods are extremely difficult to control the saccharification process in order to increase the purity of maltose or maltitol, are economically disadvantageous due to the use of a special enzyme, and have problems such as chromatographic fractionation. There are inconveniences in industrial implementation, such as the inclusion of difficult and time-consuming steps. Subsequently, the inventors focused on the fact that these inconveniences occurred due to the fact that the purity of maltose or maltitol was extremely high to facilitate pulverization, and an improvement method thereof was examined. As a result, attempts have been made to reduce the amount of oligosaccharides or oligosaccharide alcohols, even if the monosaccharides or monosaccharide reductants are slightly increased. The method of improvement is described in JP-B-57-3356, JP-B-56-28153, JP-B-56-28154, JP-A-63-101355 and JP-A-63-101356. There are some methods, and we are seeing some results. The essential point of the above-mentioned improvement method is that an enzyme having maltotriose-degrading activity / maltose-degrading activity ≧ 2.5 is allowed to act upon saccharification. A special combination was used to prepare an easily powdered sugar solution having a specific composition. (Problems to be Solved by the Invention) However, the conventional method still has problems, and cannot be said to be a sufficiently advantageous method as a method for producing maltose or maltitol which is easily powdered industrially. . For example, the above-mentioned method has a disadvantage that it is difficult to obtain an enzyme because a special enzyme is used as an enzyme for saccharification. Furthermore, since maltose is also hydrolyzed in a relatively large amount during saccharification, the glucose content increases and the maltose yield does not increase. Therefore, liquefaction is stopped at around dextrose equivalent (DE) 1 among the various conditions disclosed. Saccharification to produce high-purity maltose. That is, it was an extremely difficult method to stop liquefaction before and after DE1. In addition, the above-mentioned method has a problem that the steps are relatively long, and when maltose or maltitol with high purity is to be obtained, relatively accurate conditions such as chromatographic separation are required. . For this reason, the conventional method is not sufficient as a method for industrially advantageously producing maltose and its reduced product, which are easily powdered and contain few oligosaccharides. Under the circumstances described above, there has been a strong demand for the development of an easier and more advantageous method for industrially producing maltose having a low oligosaccharide and its reduced product. (Means for Solving the Problems) In order to solve the above-mentioned various problems, the present inventors have conducted intensive studies, and as a result, the maltogenic α-amylase of the Bacillus stearothermophilus gene has been obtained. The encoded portion is inserted into a plasmid, and a maltogenic α-amylase (hereinafter, sometimes simply referred to as a maltogenic amylase) produced by incorporating it into Bacillus subtilis is used for saccharification of starch liquefaction. By using the specific method, maltose having a small amount of oligosaccharides and a reduced product thereof, which can be easily pulverized, was successfully produced in a simpler process than the conventional method, and the present invention was completed. . Hereinafter, the contents of the present invention will be described in detail. An object of the present invention is to produce maltose having a low oligosaccharide content and its reduced product, which is easy to pulverize, makes it possible to use economically advantageous above-ground starch by using an easily available enzyme in a simple process. It is to provide a way to do it. That is, the first invention is a first step in which a liquefying enzyme is allowed to act on an aqueous starch solution having a concentration of 5 to 40% by weight to liquefy and deactivate the liquefying enzyme with a dextrose equivalent of 15 or less. A second step in which β-amylase and pullulanase and / or isoamylase are allowed to act on the product, or at the same time or within 36 hours after the start of the action, maltogenic amylase is allowed to act to saccharify, and after the start of the second step, 1 to 48 hours later Liquefied enzyme is added to 1-2 per 1g of solid substrate.
0 units are added to further saccharify, the maltose content of the saccharified product is 75 to 90% by weight in the solid content, and the content of the oligosaccharide contained in the saccharified product is in the range of 7 or less when calculated by the following formula. The third step of saccharification, Consists of The second invention is a first step in which a liquefying enzyme is allowed to act on an aqueous starch solution having a concentration of 5 to 40% by weight to liquefy and deactivate the liquefying enzyme with a dextrose equivalent of 15 or less. A second step in which β-amylase and pullulanase and / or isoamylase are allowed to act on the product, or at the same time or within 36 hours after the start of the action, maltogenic amylase is allowed to act to saccharify, and after the start of the second step, 1 to 48 hours later Liquefied enzyme is added to 1-2 per 1g of solid substrate.
0 units are added to further saccharify, the maltose content of the saccharified product is 75 to 90% by weight in the solid content, and the content of the oligosaccharide contained in the saccharified product is in the range of 7 or less when calculated by the following formula. The third step of saccharification, A fourth step of reducing the saccharified product obtained in the third step. Maltose or a reduced product thereof having a low oligosaccharide obtained through the steps of the above inventions is subjected to purification, concentration, chromatographic separation, crystallization (solidification), drying, powdering, etc. by a known method. This makes it possible to easily prepare powdery or crystalline maltose or maltitol, and further to prepare highly pure maltose or maltitol. Hereinafter, the content of the present invention will be described in more detail. First, the content of the first invention will be described. In the present invention, ground starch and underground starch can be used irrespective of whether they are ground starch or ground starch. In particular, ground starch which is conventionally inconvenient in producing powdery or high-purity maltose and its reduced product can be advantageously used. This is one of the advantages of the present invention. In practicing the present invention, the composition of amylose and amylopectin in the starch does not need to be particularly considered, and concrete examples of usable starch include corn starch, wheat starch, barley starch, and ground starch such as starch. In addition, various underground starches can be mentioned. When liquefying these starches, when ground starch is used as a raw material, the substrate concentration during liquefaction is preferably adjusted to 10 to 30%, and the pH is adjusted to 6.0 to 6.8 in order to prevent aging of the liquefied liquid. Liquefaction enzyme It is desirable to liquefy using a liquefaction enzyme such as Termamil (registered trademark) of Novo, and to inactivate the liquefaction enzyme with a dextrose equivalent of 15 or less, more preferably 3 to 13. Next, β-amylase, pullulanase and / or isoamylase and maltogenic amylase are allowed to act on the liquefied product simultaneously with β-amylase and pullulanase and / or isoamylase within ~ 36 hours, and saccharification is performed. Preferred conditions are substrate concentration 5-40% by weight, pH
5.3, temperature is about 55 ° C. 1 to 48 hours after the start of saccharification, 1 to 20 units of liquefying enzyme is added per 1 g of solid substrate to further saccharify. By this operation, mainly oligosaccharides of four or more saccharides are hydrolyzed to form maltose and trisaccharide. And, if necessary, a sugar composition that promotes the further development of the effects of the subsequent steps, thereby improving the filterability. The enzymes used at this time were β-amylase # 1500 manufactured by Nagase Sangyo Co., Ltd. and SPEZYME (registered trademark) BB manufactured by Finsugar Co., Ltd.
A 1500 and the like, and among them, β-amylase derived from soybean has advantageous properties in practicing the present invention, and as pullulanase, Promozyme from Novo or Pullulanase Amano CKL from Amano Pharmaceutical Co., Ltd. And the like are highly versatile, and can be advantageously used because they are commercially available and the properties of enzymes. On the other hand, as a maltogenic amylase produced by the genetic modification, there is a maltogenase (Maltogenase; registered trademark) of Novo. The preferred ratio of the amount of the enzyme to be used in the saccharification is, for example, 1 to 20 units of Novo's maltogenase produced by the genetic modification (the activity of this maltogenase will be described using a maltogenic amylase novo unit). ) When used, β-amylase is 10-30 units and pullulanase is 0.6-2.0 units. This saccharification step is carried out until the maltose purity reaches equilibrium (usually 24 to 72).
Time). The liquefied enzyme used in the third step can be used with either non-thermostable or thermostable, but non-thermostable liquefied enzyme is more effective in practicing the present invention. According to the above-mentioned steps, the saccharification can be performed in the range of a numerical value of 7 or less when the maltose purity is 75 to 90% by weight and the content of the oligosaccharide contained in the saccharified product is calculated by the following equation. The saccharified product obtained by this step has a relatively high maltose purity among the currently marketed maltose-based products, and its component composition is low in oligosaccharide content of trisaccharide or more. The viscosity is lower than that of similar products on the market, and when crystallized, the crystal growth rate of maltose is high, so that it is easy to directly crystallize and powder by a known method. Further, even when the purity is increased by subjecting it to a crystallization or chromatographic separation step, high-purity maltose can be produced more advantageously than a product produced by a conventional method. As a method for producing powdered maltose, for example, various methods such as a honey separation method, a spray granulation method, a fluid granulation method, a block pulverization method, or a combination thereof can be adopted. If necessary, to increase the maltose purity, various ion exchangers such as ion-exchange resin, ion-exchange fiber, zeolite, etc. are converted to alkali metal type and chromatographed. It is possible to adopt a method for converting the data to a new one. Next, the step of the second invention will be described. The second invention is to improve the maltose purity of the saccharified product obtained in the first invention as it is or as necessary by the above various means, and then by a known batch or continuous method itself, Hydrogenation is carried out in the presence of a nickel-based or noble metal-based reduction catalyst to form a maltitol-based sugar alcohol. The hydrogenation condition may be any condition as long as maltose does not decompose. Usually, the concentration of the sugar solution is 40 to 60% by weight, and the hydrogen pressure is 20 kg / cm ² or more, more preferably. 5
It is carried out at a temperature of 100-150 ° C. at 0-200 kg / cm ² . Maltitol with a low oligosaccharide content obtained by this process has a relatively high maltitol purity among the currently marketed maltitol-based products, and its component composition is trisaccharide Since the above oligosaccharide alcohol content is low, the viscosity is lower than that of similar products on the market, and when crystallized, the crystal growth rate of maltitol is high. Is easy. Thereafter, if necessary, the purity of maltitol can be easily further increased by a known method such as a chromatographic separation method or crystallization separation. (Example) Next, the content of the present invention will be described more specifically with reference to examples, but the present invention is not limited to the following examples. Example-1 First step (liquefaction step) Corn starch was adjusted to a concentration of 30% and pH 6.2, and a heat-resistant liquefying enzyme [Nagase Sangyo Co., Ltd.
20u / g substrate solid content (hereinafter abbreviated as DS) was added, and the mixture was liquefied at 105 ° C by a conventional method. The liquefaction was stopped by heating at DE 6.5. Second step Next, after adjusting the temperature to 55 ° C. and the pH to 5.3, 10u / g DS β-manufactured by Nagase & Co., Ltd.
-Amylase # 1500 and 200L of Promozyme TM from Novo of 0.67u / gDS were added to proceed the saccharification reaction. Six hours after the start of the second step, 6.5 u / g DS of Novart's maltogenase was added. Third step Six hours after the start of the second step, 20 u / g DS of the above-mentioned spinase PN-4 was added, and the saccharification reaction was continued for another 66 hours. The result of measurement of the sugar composition after completion of the third step by high performance liquid chromatography was as follows. Monosaccharide 7.0% Disaccharide 88.2% Trisaccharide 1.0% Oligosaccharide with more than tetrasaccharide 3.8% Example-2 Purification of 50% concentration by decolorizing, desalting and concentrating the saccharified product obtained in Example-1 according to a conventional method. 20 kg of the sugar solution and 200 g of Raney nickel catalyst were charged into an autoclave having an internal volume of 25 l, and the mixture was stirred at 120 ° C. for 2 hours while maintaining the hydrogen pressure at 150 to 120 kg / cm ² , and hydrogenated. After the obtained liquid was separated from the catalyst, the result of analysis by high performance liquid chromatography through a column of granular activated carbon was as follows. Sorbitol 7.4% Maltitol 88.1% Trisaccharide or higher oligosaccharide alcohol 4.5% After purification, the resulting reduced product is concentrated to 75%, cooled to 10 ° C, 3% by weight of seed crystals are added and mixed. For 15 hours, a mass kit was prepared and spray-crystallized at a blowing temperature of 80 ° C. with a spray dryer to obtain crystalline powder maltitol. Comparative Example-1 For comparison, a commercially available maltose solution (sugar composition = monosaccharide 1.4%, disaccharide 90.0%, trisaccharide 7.0)
%, 1.6% oligosaccharide or more) to obtain a reduced product having the following composition. Sorbitol 1.5% Maltitol 89.6% Trisaccharide or higher oligosaccharide alcohol 8.9% After the purification, concentration and cooling as in Example-2, mass kit preparation was performed.
After 5 hours, no spray-dryable mass kit was obtained. (Effect of the Invention) As described above, by carrying out the present invention, an oligosaccharide having a composition which can be easily powdered with a relatively short process, easy process control, and economically advantageous can be obtained. It becomes possible to produce less maltose and its reduced products.

Claims

Claims 1. A liquefying enzyme is allowed to act on a ground starch aqueous solution having a concentration of 10 to 30 % by weight to liquefy.
A first step in which the liquefaction enzyme is inactivated with a dextrose equivalent of 15 or less, simultaneously with the action of β-amylase and pullulanase and / or isoamylase on the liquefied product obtained in the above step, or within 36 hours after the start of the action; The maltogenic α-amylase-encoded portion of the Bacillus stearothermophilus gene is inserted into a plasmid, and
Maltogenic-α produced by incorporation into Subtilis (Bacil-lus subtilis)
-A second step of saccharification by the action of amylase; 1 to 48 hours after the start of the second step, the liquefied enzyme is added to the saccharified enzyme in an amount of 1 to 2 per gram of solid substrate.
0 units are added to further saccharify, the maltose content of the saccharified product is 75 to 90% by weight in the solid content, and the content of the oligosaccharide contained in the saccharified product is in the range of 7 or less when calculated by the following formula. The third step of saccharification, A method for producing maltose having a low content of oligosaccharides, wherein the three steps are sequentially performed. 2. Liquefaction by acting a liquefaction enzyme on a ground starch aqueous solution having a concentration of 10 to 30 % by weight,
A first step in which the liquefaction enzyme is inactivated with a dextrose equivalent of 15 or less, simultaneously with the action of β-amylase and pullulanase and / or isoamylase on the liquefied product obtained in the above step, or within 36 hours after the start of the action; The maltogenic α-amylase-encoded portion of the Bacillus stearothermophilus gene is inserted into a plasmid, and
Maltogenic-α produced by incorporation into Subtilis (Bacil-lus subtilis)
-A second step of saccharifying by the action of an amylase; 1 to 48 hours after the start of the second step, the liquefied enzyme is added in an amount of 1 to 2 per gram of solid substrate.
0 units are added to further saccharify, the maltose content of the saccharified product is 75 to 90% by weight in the solid content, and the content of the oligosaccharide contained in the saccharified product is in the range of 7 or less when calculated by the following formula. The third step of saccharification, A fourth step of reducing the saccharified product obtained in the third step; and a method for producing maltitol having a small amount of oligosaccharide alcohol, wherein the four steps are sequentially performed.