JP6169896B2 - Heavy metal adsorbent and heavy metal recovery method - Google Patents
Heavy metal adsorbent and heavy metal recovery method Download PDFInfo
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Description
本発明は、金属の吸着剤、さらに詳しくは担子菌の死菌体又はこれを含有する廃菌床を利用した重金属吸着剤、及びこれらを使用する重金属回収方法等に関する。 The present invention relates to a metal adsorbent, and more particularly to a heavy metal adsorbent using a dead cell of a basidiomycete or a waste fungus bed containing the same, a heavy metal recovery method using the same, and the like.
廃水中に存在する金属類を微生物の細胞表面などに吸着させるバイオソープション技術は、有害金属の除去、希少金属のリサイクルなどに重要であり、各種の材料、化学物質及び多様な微生物の細胞を吸着剤として利用することが研究されている。 Biosorption technology that adsorbs metals present in wastewater to the cell surface of microorganisms is important for removing harmful metals, recycling rare metals, and so on. Various materials, chemical substances, and cells of various microorganisms are used. The use as an adsorbent has been studied.
微生物の細胞を利用した吸着剤として細菌、カビ、酵母が研究されている。しかし、微生物を利用した金属吸着剤の実用化に向けては菌体の確保が大きな課題となる。カビは、成長過程で大量の胞子を産生する菌種が多いため、毒性の少ない菌種であってもアレルギーなどの問題を引き起こす可能性がある。また、細菌や酵母の多くは単細胞で、細胞の寸法が小さく、吸着後の回収が難しいという問題点がある。 Bacteria, molds, and yeasts have been studied as adsorbents using microbial cells. However, securing the bacterial cells is a major issue for the practical application of metal adsorbents using microorganisms. Molds often produce large amounts of spores during the growth process, and even less toxic species can cause problems such as allergies. In addition, many of bacteria and yeast are single cells, and have a problem that the cell size is small and recovery after adsorption is difficult.
カビと同様に真菌である担子菌については、担子菌（レイシ、Ganoderma lucidum）の成長に伴って菌体内に金属を蓄積させる方法が報告されている（特許文献１）。しかし、微生物の成長に伴って菌体内外に金属を蓄積させる方法は、生きた細胞を利用するため、微生物の成長を促すための栄養を定期的に与える必要があり、吸着に時間を要する。また、成長阻害が生じるような高濃度金属廃水には適用できないこと、細胞内に蓄積された金属の回収が困難であることなどの問題がある。 As for basidiomycetes, which are fungi as well as molds, a method has been reported in which metals are accumulated in the fungus body as the basidiomycete ( Ganoderma lucidum ) grows (Patent Document 1). However, the method of accumulating metals inside and outside the cells with the growth of microorganisms uses living cells, so it is necessary to periodically provide nutrients for promoting the growth of microorganisms, and it takes time for adsorption. In addition, there are problems that it cannot be applied to high-concentration metal wastewater that causes growth inhibition, and that it is difficult to recover metal accumulated in cells.
担子菌を材料として製造した吸着剤については、ロシア特許第２０７３０１５号公報（特許文献２）及び特表２００２−５０６９７９号公報（特許文献３）において細胞壁由来のキチン−グルカン−メラミン複合体を含む物質を放射性核種等の吸着剤として用いることが記載されている。これらの文献に記載された吸着剤では、Coltricia、Coriolus、Daedaleaなどの担子菌を使用した吸着剤に放射性核種等及び金属を吸着させたことが記載されているが、吸着後の金属の回収については記載されていない。 About the adsorbent manufactured using basidiomycetes as a material, a substance containing a chitin-glucan-melamine complex derived from a cell wall in Russian Patent No. 2073015 (Patent Document 2) and Japanese Translation of PCT International Publication No. 2002-506969 (Patent Document 3) Is used as an adsorbent for radionuclides and the like. The adsorbents described in these documents describe that radionuclides and metals were adsorbed to adsorbents using basidiomycetes such as Coltricia , Coriolus , Daedalea, etc. Is not listed.
そこで、本発明は、液体中の金属、特に廃水中の重金属を迅速に吸着した後、効率よく回収できる吸着剤及びそれを用いた重金属の回収方法を提供することを目的とする。また、本発明は、安全性が高く、入手が容易な材料を使用することができ、製造及び使用が簡便で、経済的にも有利な吸着剤及び重金属の回収方法を提供することを目的とする。 Accordingly, an object of the present invention is to provide an adsorbent that can be efficiently recovered after rapidly adsorbing a metal in a liquid, particularly heavy metal in wastewater, and a method for recovering heavy metal using the adsorbent. Another object of the present invention is to provide an adsorbent and a method for recovering heavy metals that can use materials that are highly safe and easily available, that are simple to manufacture and use, and that are economically advantageous. To do.
本発明は、
〔１〕 担子菌の死菌体を含有する重金属吸着剤；
〔２〕 前記担子菌の死菌体が、乾燥処理、アルカリ処理、オゾン処理、又はアルカリ処理及びオゾン処理されたものである、〔１〕記載の重金属吸着剤；
〔３〕 前記担子菌の死菌体が、廃菌床に含まれている、〔１〕記載の重金属吸着剤；
〔４〕 粉体状である、〔１〕記載の重金属吸着剤；
〔５〕 前記担子菌が、ハラタケ目（Agaricales）又はチョレマイタケ目（Polyporales）に属する担子菌である、〔１〕記載の重金属吸着剤；
〔６〕 前記担子菌が、Pleurotus属、Hypsizygus属、Lentinula属、Flammulina属及びGrifola属からなる群より選択された一種以上の担子菌である、〔１〕記載の重金属吸着剤；
〔７〕 前記担子菌が、エリンギ（Pleurotus eryngii）、ブナシメジ（Hypsizygus marmoreus）、シイタケ（Lentinula edodes）、エノキタケ（Flammulina velutipes）及びマイタケ（Grifola frondosa）からなる群より選択された一種以上の担子菌である、〔１〕記載の重金属吸着剤；
〔８〕 重金属の回収方法であって、重金属を含有させた〔１〕〜〔７〕のいずれか１項記載の重金属吸着剤を、酸性液体と接触させることにより、前記酸性液体中に重金属を回収する工程を含む、方法；
〔９〕 前記酸性液体が、ｐＨ４以下の塩酸又は硫酸である、〔８〕記載の方法；
〔１０〕 回収される重金属が、ニッケル、コバルト及びセシウムからなる群より選択される一以上の重金属である、〔８〕記載の方法；
〔１１〕 担子菌の死菌体を含有する重金属吸着剤の製造方法であって、担子菌又は担子菌を含む廃菌床を、オゾン処理した後にアルカリ処理をする工程を含む、方法
を提供する。
The present invention
[1] Heavy metal adsorbent containing dead basidiomycetes;
[2] The heavy metal adsorbent according to [1], wherein the dead cells of the basidiomycete are dried, alkali-treated, ozone-treated, or alkali-treated and ozone-treated;
[3] The heavy metal adsorbent according to [1], wherein the dead cells of the basidiomycete are contained in a waste fungus bed;
[4] The heavy metal adsorbent according to [1], which is in powder form;
[5] The heavy metal adsorbent according to [1], wherein the basidiomycete is a basidiomycete belonging to the order Agaricales or Polyporales ;
[6] The heavy metal adsorbent according to [1], wherein the basidiomycete is one or more basidiomycetes selected from the group consisting of Pleurotus , Hypsizygus , Lentinula , Flammulina and Grifola .
[7] The basidiomycete is one or more basidiomycetes selected from the group consisting of eryngii ( Pleurotus eryngii ), beech shimeji ( Hypsizygus marmoreus ), shiitake ( Lentinula edodes ), enokitake ( Flammmulina velutipes ) and maitake ( Grifola frondosa ) A heavy metal adsorbent according to [1],
[8] A method for recovering heavy metal, wherein the heavy metal adsorbent according to any one of [1] to [7] containing heavy metal is brought into contact with the acidic liquid, whereby the heavy metal is contained in the acidic liquid. A method comprising the step of recovering;
[9] The method according to [8], wherein the acidic liquid is hydrochloric acid or sulfuric acid having a pH of 4 or less;
[10] The method according to [8], wherein the recovered heavy metal is one or more heavy metals selected from the group consisting of nickel, cobalt and cesium;
[11] A method for producing a heavy metal adsorbent containing dead bacilli of basidiomycetes, comprising a step of performing an alkali treatment after ozone treatment of basidiomycetes or a waste fungus bed containing basidiomycetes. .
本発明によれば、重金属吸着速度が速く、吸着した重金属の脱離回収が容易な重金属吸着剤が提供される。本発明の吸着剤は、担子菌、及び廃菌床等の担子菌含有物における担子菌の、死細胞を利用するので、細胞の成長を必要としない。したがって、細胞の成長のための栄養分添加の必要がなく、また、重金属による担子菌の成長阻害を考慮する必要がない。そのため、本発明によれば、簡便な方法で、低コストで高濃度の廃水からの重金属の回収が可能である。 ADVANTAGE OF THE INVENTION According to this invention, the heavy metal adsorption | suction rate is quick and the heavy metal adsorption agent with which the adsorption | suction recovery of the adsorbed heavy metal is easy is provided. Since the adsorbent of the present invention uses dead cells of basidiomycetes and basidiomycetes in a basidiomycete-containing product such as a waste fungus bed, cell growth is not required. Therefore, it is not necessary to add nutrients for cell growth, and it is not necessary to consider the inhibition of basidiomycete growth by heavy metals. Therefore, according to the present invention, it is possible to recover heavy metals from wastewater having a high concentration at a low cost by a simple method.
また、本発明の吸着剤は重金属の吸収効率が高く、さらに、吸着した重金属を簡便な操作で９０％以上もの高い回収率で容易に回収することができる。そのため、廃液から有害な重金属を除去する目的のほか、希少又は高価な重金属を吸着後に回収して、再利用することが可能となる。 Further, the adsorbent of the present invention has a high heavy metal absorption efficiency, and can easily recover the adsorbed heavy metal with a high recovery rate of 90% or more by a simple operation. Therefore, in addition to the purpose of removing harmful heavy metals from the waste liquid, it is possible to collect rare or expensive heavy metals after adsorption and reuse them.
さらに、担子菌の菌体はある程度成長すると目視で確認できる程度の大きさがあるため、菌体を回収する際などの取り扱いが容易である。また、本発明において使用される担子菌としては、食用の担子菌を利用することができ、製造中及び使用中の安全性が高い。 Furthermore, since the basidiomycetous cells have a size that can be visually confirmed when they grow to some extent, they are easy to handle when collecting the cells. Moreover, as a basidiomycete used in this invention, an edible basidiomycete can be utilized and the safety | security during manufacture and use is high.
特に、食用の担子菌は食用キノコとしておがくず等による菌床栽培が行われており、キノコを収穫した後には多量の菌糸を含むおがくず等が廃菌床として残る。これを利用することにより、通常は廃棄される廃菌床の有効利用にもなり、経済的にもさらに有利に吸着剤を製造することができる。 In particular, edible basidiomycetes are cultivated by sawdust etc. as edible mushrooms, and sawdust containing a large amount of hyphae remains as a waste fungus bed after harvesting mushrooms. By utilizing this, it becomes effective utilization of the waste microbial bed normally discarded, and an adsorbent can be manufactured further advantageously economically.
本発明の吸着剤は、担子菌、又は廃菌床等の担子菌含有物中の担子菌の、死菌体を含むことを特徴とする。本発明に使用される担子菌種としては、特に制限はないが、子実体を形成する担子菌、特にハラタケ目（Agaricales）又はチョレマイタケ目（Polyporales）の担子菌を使用することができる。 The adsorbent of the present invention is characterized by containing dead cells of basidiomycetes or basidiomycetes in a basidiomycete-containing material such as a waste fungus bed. The basidiomycete species used in the present invention is not particularly limited, and basidiomycetes that form fruit bodies, particularly basidiomycetes ( Agaricales ) or choremaitake ( Polyporales ) basidiomycetes can be used.
本発明の担子菌として、食用キノコとして栽培されている種類の担子菌、たとえばPleurotus属、Hypsizygus属、Lentinula属、Flammulina属、Grifola属の担子菌（代表的なものとして、具体的にはPleurotus eryngii（エリンギ）、Hypsizygus marmoreus（ブナシメジ）、Lentinula edodes（シイタケ）、Flammulina velutipes（エノキタケ）、Grifola frondosa（マイタケ）などが挙げられる）などを使用することができる。このような食用キノコとして栽培されている担子菌は、一般に成長が早く、安全性が高いうえ、容易に入手可能である点で有利である。特に、このような食用として菌床栽培されている担子菌の、可食部を収穫した後の廃菌床は、一回又は数回使用された後に廃棄物として処分されることが多く、大量に入手可能であり、廃物の有効利用となる点で好ましい。 As the basidiomycetes of the present invention, the types of basidiomycetes cultivated as edible mushrooms, such as the basidiomycetes of the genera Pleurotus , Hypsizygus , Lentinula , Flammulina , Grifola (typically, specifically, Pleurotus eryngii (Eringi), Hypsizygus marmoreus (Bunashimeji), Lentinula edodes (Shiitake), Flammulina velutipes ( Enotake ), Grifola frondosa (Maitake), etc.) can be used. Basidiomycetes cultivated as such edible mushrooms are advantageous in that they are generally fast growing, high in safety, and easily available. In particular, the basidiomycetes that have been cultivated for edible fungi, the waste beds after harvesting the edible part are often disposed of as waste after being used once or several times. It is preferable in that it can be easily obtained and used effectively.
廃菌床とは、担子菌を栽培した後の菌床であり、栽培対象の担子菌（及び場合によってはその他の微生物）と、菌床を構成する成分との混合物である。菌床を構成する成分は、栽培対象の菌種によって異なるが、一般に、おがくず、コーンコブなどの植物細胞を含む培養基材、米糠、フスマなどの栄養剤、各種の微量添加剤などを含む。本発明に使用される廃菌床は、担子菌が含まれていればよく、これらの菌床構成成分のすべてを含むことは必要としない。 The waste fungus bed is a fungus bed after cultivation of basidiomycetes, and is a mixture of basidiomycetes to be cultivated (and other microorganisms in some cases) and components constituting the fungus bed. The components constituting the fungus bed vary depending on the cultivated species, but generally include a culture substrate containing plant cells such as sawdust and corn cob, nutrients such as rice bran and bran, various trace additives, and the like. The waste microbial bed used in the present invention only needs to contain basidiomycetes, and does not need to contain all of these microbial bed components.
本発明の吸着剤においては、担子菌の死菌体を利用する。死菌体は、担子菌又は廃菌床などの担子菌含有物を乾燥させることにより容易に得ることができるが、オートクレーブ処理など、公知のいずれの手段によって得てもよい。 In the adsorbent of the present invention, killed cells of basidiomycetes are used. Although dead cells can be easily obtained by drying basidiomycetes-containing materials such as basidiomycetes or waste fungus beds, they may be obtained by any known means such as autoclaving.
本発明の吸着剤は、死菌体を含む担子菌又は担子菌含有物を、乾燥させた後、粉砕することにより、製造することができる。重金属の吸着量及び速度を向上させるためには、吸着剤の表面積を増大させることが望ましいと考えられる。したがって、本発明の吸着剤は、粉体状であることが好ましい。粉体は、本明細書においては、特定の粒子サイズを意図するものではなく、細かい粒状の固体の集合体を意味するものであり、粉粒体、微粒子、（微）粉末などの用語と互換的なものとして使用される。粉砕は、公知の任意の機械的方法などを用いて行うことができ、さらに必要に応じてふるい分けを行うことなどにより、粒子サイズを所望の範囲にそろえることができる。 The adsorbent of the present invention can be produced by drying and then crushing basidiomycetes containing dead cells or basidiomycetes-containing materials. In order to improve the adsorption amount and speed of heavy metals, it is considered desirable to increase the surface area of the adsorbent. Therefore, the adsorbent of the present invention is preferably in the form of powder. In the present specification, the powder is not intended to be a specific particle size, but means an aggregate of fine granular solids, and is interchangeable with terms such as powder, fine particles, and (fine) powder. Used as a standard. The pulverization can be performed using any known mechanical method, and further, the particle size can be adjusted to a desired range by performing sieving as necessary.
本発明の吸着剤は、死菌体をそのまま利用することもできるが、これをさらにアルカリ処理、オゾン処理などの処理をすることにより、吸着効率の高い吸着剤とすることができる。具体的には、たとえば、アルカリ処理は、死菌体をアルカリ性の溶液（たとえば水酸化ナトリウムなどの溶液）と接触させることで行うことができ、その後、菌体を蒸留水、イオン交換水及び超純水などで洗浄することにより中性のｐＨに戻し、再度乾燥（通常、約７０℃以下）させることができる。アルカリ処理及び洗浄は、通常、約４℃〜約１００℃で約５分間〜約６０分間程度行うことができるが、特に制限はない。本発明におけるアルカリ処理は、室温に近い温度（たとえば約１０℃〜約４０℃）で簡便に行うことができる。当業者は適宜条件を選択することができる。 The adsorbent of the present invention can use dead cells as they are, but can be made into an adsorbent with high adsorption efficiency by further treatment with alkali treatment, ozone treatment or the like. Specifically, for example, the alkali treatment can be performed by bringing dead cells into contact with an alkaline solution (for example, a solution such as sodium hydroxide), and then the cells are subjected to distilled water, ion-exchanged water, and ultrapure water. It can be returned to neutral pH by washing with pure water or the like and dried again (usually about 70 ° C. or lower). The alkali treatment and washing can be usually performed at about 4 ° C. to about 100 ° C. for about 5 minutes to about 60 minutes, but there is no particular limitation. The alkali treatment in the present invention can be easily performed at a temperature close to room temperature (for example, about 10 ° C. to about 40 ° C.). Those skilled in the art can appropriately select the conditions.
オゾン処理は、死菌体をアルカリ処理した後に、あるいはアルカリ処理を行う前に、あるいはアルカリ処理を行わずに単独で、行うことができる。オゾン処理は、オゾン発生装置などで発生させたオゾンを含む雰囲気中で、死菌体を保持することにより行うことができる。当業者は適宜条件を選択することができるが、一般的には、たとえば約４℃〜約４０℃、約５分間〜約３時間程度で処理を行うことができる。アルカリ処理を行う前にオゾン処理を行うことにより、吸着剤の金属吸着量を増大させることができる。したがって、アルカリ処理とオゾン処理とを両方行う場合は、オゾン処理を行った後にアルカリ処理を行うことが望ましい。 The ozone treatment can be carried out after the dead cells are treated with an alkali, before the treatment with an alkali, or without an alkali treatment. The ozone treatment can be performed by holding dead cells in an atmosphere containing ozone generated by an ozone generator or the like. Those skilled in the art can appropriately select the conditions, but in general, for example, the treatment can be performed at about 4 ° C. to about 40 ° C. for about 5 minutes to about 3 hours. By performing the ozone treatment before the alkali treatment, the amount of metal adsorbed by the adsorbent can be increased. Therefore, when performing both alkali treatment and ozone treatment, it is desirable to perform alkali treatment after ozone treatment.
本発明の吸着剤は、吸着すべき対象の重金属を含有する液体、たとえば廃水と接触させることにより、使用することができる。ここで、重金属とは、比重が５以上の金属を指し、クロム、コバルト、ニッケル、亜鉛、銀、金、ウランなどが挙げられる。また、セシウムのような原子量５０以上のアルカリ金属も吸着し得る。本発明の吸着剤により吸着すべき対象の金属としては、水溶液中で陽イオン化するものが適している。 The adsorbent of the present invention can be used by bringing it into contact with a liquid containing the heavy metal to be adsorbed, for example, waste water. Here, the heavy metal refers to a metal having a specific gravity of 5 or more, and includes chromium, cobalt, nickel, zinc, silver, gold, uranium and the like. Further, an alkali metal having an atomic weight of 50 or more such as cesium can also be adsorbed. As the target metal to be adsorbed by the adsorbent of the present invention, those that are cationized in an aqueous solution are suitable.
本発明の吸着剤と重金属を含む液体との接触は、吸着剤を液体中に投入し、必要に応じて攪拌することで容易に実施することができる。接触時の温度、時間などは、吸着対象の金属の種類、濃度、吸着剤の使用量などによって適宜調整することができるが、約４℃〜約６０℃、約５分〜約３時間程度とすることができる。本発明の吸着剤は室温に近い温度で非常に迅速に重金属を飽和濃度まで吸着することができる。 The contact between the adsorbent of the present invention and the liquid containing heavy metal can be easily carried out by putting the adsorbent into the liquid and stirring it as necessary. The temperature and time at the time of contact can be appropriately adjusted depending on the type and concentration of the metal to be adsorbed, the amount of adsorbent used, etc., and are about 4 ° C. to about 60 ° C., about 5 minutes to about 3 hours. can do. The adsorbent of the present invention can adsorb heavy metals to a saturated concentration very rapidly at a temperature close to room temperature.
吸着した重金属は、吸着剤を酸性の溶液と接触させることにより、容易に吸着剤から溶液中に脱離させることができる。酸性の溶液としては、ｐＨ４以下のものが好ましく、特にｐＨ１〜３のものが好ましい。酸性の溶液として、具体的には、塩酸、硫酸、硝酸、乳酸などが挙げられるが、塩酸又は硫酸が好ましい。 The adsorbed heavy metal can be easily desorbed from the adsorbent into the solution by bringing the adsorbent into contact with an acidic solution. As the acidic solution, those having a pH of 4 or less are preferred, and those having a pH of 1 to 3 are particularly preferred. Specific examples of the acidic solution include hydrochloric acid, sulfuric acid, nitric acid, and lactic acid, but hydrochloric acid or sulfuric acid is preferable.
酸性溶液と吸着剤との接触は、吸着剤を酸性溶液中に投入し、必要に応じて攪拌することによって容易に実施することができる。吸着剤からの重金属の脱離に要する時間は、酸性溶液のｐＨ、酸の種類、吸着した金属の種類、吸着量、菌種などによって変動し得るが、概ね約４℃〜約１００℃、約５分〜約３時間程度とすることができる。本発明の吸着剤は室温に近い温度で非常に迅速に重金属を酸性溶液中に放出することができる。 The contact between the acidic solution and the adsorbent can be easily carried out by putting the adsorbent into the acidic solution and stirring as necessary. The time required for desorption of heavy metal from the adsorbent may vary depending on the pH of the acidic solution, the type of acid, the type of adsorbed metal, the amount of adsorption, the bacterial species, etc., but is generally about 4 ° C. to about 100 ° C., about It can be about 5 minutes to about 3 hours. The adsorbent of the present invention can release heavy metals into an acidic solution very rapidly at temperatures close to room temperature.
重金属を含む酸性溶液は、回収した金属の種類、濃度及び単一金属溶液か複合金属溶液かなどによって選択した公知の方法でそれぞれ適宜処理することにより、目的の金属をさらに濃縮したり固体として取り出したりすることができる。 The acidic solution containing heavy metals is appropriately treated by a known method selected according to the type and concentration of the recovered metal and whether it is a single metal solution or a composite metal solution, thereby further concentrating the target metal or taking it out as a solid. Can be.
１．担子菌を含む吸着剤の製造
Pleurotus eryngii（エリンギ）、Hypsizygus marmoreus（ブナシメジ）、Lentinula edodes（シイタケ）、Flammulina velutipes（エノキタケ）及びGrifola frondosa（マイタケ）の担子菌類５菌種の子実体を用いた。
1. Production of adsorbents containing basidiomycetes
The fruiting bodies of 5 basidiomycetous species of Pleurotus eryngii (Eringi), Hypsizygus marmoreus (Bunashimeji), Lentinula edodes (Shiitake), Flammulina velutipes (Enokitake) and Grifola frondosa (Maitake) were used.
子実体を、乾燥機で６０℃、３日間乾燥させ、それを粉砕機で２０秒間粉砕し、目開き３５５μmのふるい通過分をさらに６０℃、２４時間乾燥させた（「無処理担子菌」とする）。
無処理担子菌１０gを０．２M 水酸化ナトリウム溶液１Lに投入し、室温で３０分間マグネチックスターラーを用いて撹拌することにより、アルカリ処理を行った。この菌体を、洗浄液のｐＨが中性域に達するまでイオン交換水を用いて洗浄した。その後、遠心分離及び吸引ろ過によって菌体を回収し、乾燥器で６０℃、２日間乾燥させ、それを粉砕機で粉砕し、目開き３５５μmのふるい通過分をさらに６０℃、２４時間乾燥させた（「アルカリ処理担子菌」とする）。
The fruiting bodies were dried at 60 ° C. for 3 days with a dryer, pulverized with a pulverizer for 20 seconds, and the sieve passage having an opening of 355 μm was further dried at 60 ° C. for 24 hours (“Untreated basidiomycetes”) To do).
10 g of untreated basidiomycetes were put into 1 L of 0.2M sodium hydroxide solution and stirred with a magnetic stirrer at room temperature for 30 minutes for alkali treatment. The cells were washed with ion exchange water until the pH of the washing solution reached a neutral range. Thereafter, the cells were collected by centrifugation and suction filtration, dried at 60 ° C. for 2 days with a dryer, pulverized with a pulverizer, and further dried through a sieve having an opening of 355 μm at 60 ° C. for 24 hours. (Referred to as “alkali-treated basidiomycetes”).
アルカリ処理担子菌の一部について、さらにオゾン処理を行った。１０Lのガラス製デシケータ内に担子菌０．１gを静置し、オゾン発生装置（日本オゾン株式会社製、ＯＮ−３−２型）を用いて発生させたオゾンをデシケータ内に導入した。オゾンの発生量は３．０mg/minでオゾンの導入は約１時間とした（「オゾン処理担子菌」とする）。 Ozone treatment was further performed on some of the alkali-treated basidiomycetes. 0.1 g of basidiomycetes were allowed to stand in a 10 L glass desiccator, and ozone generated using an ozone generator (manufactured by Nippon Ozone Co., Ltd., ON-3-2 type) was introduced into the desiccator. The amount of ozone generated was 3.0 mg / min, and ozone was introduced for about 1 hour (referred to as “ozone-treated basidiomycete”).
２．液体からの金属の吸着実験
ニッケル（Ｎｉ2+）、コバルト（Ｃｏ2+）、リチウム（Ｌｉ+）、ホウ素（［Ｂ（ＯＨ）４］-）、又はセシウム（Ｃｓ+）を含む金属溶液は、それぞれＮｉ（ＮＯ3）2・６Ｈ2Ｏ（関東化学(株)）、Ｃｏ（ＮＯ3）2・６Ｈ2Ｏ（関東化学(株)）、ＬｉＣｌ（関東化学(株)）、Ｈ3ＢＯ3（関東化学(株)又はＣｓＣｌ（関東化学(株)）を１ppm〜５０ppmになるように蒸留水に溶解して調製した。各金属溶液のｐＨは、０．１M 水酸化ナトリウム及び／又は０．１M 硝酸（ＨＮＯ3）を使用してｐＨ６に調整した。
2. Experiment of metal adsorption from liquid Metal solution containing nickel (Ni 2+ ), cobalt (Co 2+ ), lithium (Li + ), boron ([B (OH) 4] − ), or cesium (Cs + ) , Ni (NO 3 ) 2 · 6H 2 O (Kanto Chemical Co., Inc.), Co (NO 3 ) 2 · 6H 2 O (Kanto Chemical Co., Ltd.), LiCl (Kanto Chemical Co., Ltd.), H 3 BO 3 (Kanto Chemical Co., Ltd.) or CsCl (Kanto Chemical Co., Ltd.) was prepared by dissolving in distilled water to 1 ppm to 50 ppm. The pH of each metal solution was 0.1 M sodium hydroxide and / or 0 Adjusted to pH 6 using 1M nitric acid (HNO 3 ).
２．１ 吸着量の測定
１ppm〜５０ppmの金属溶液１Lに対して無処理担子菌及びアルカリ処理担子菌又はオゾン処理担子菌を１gの割合で添加した。混合液は３０℃に設定した恒温器内でマグネチックスターラーを用いて攪拌した。吸着処理後の溶液は随時採取し、０．４５μmのアクリルスチロール製メンブランフィルター（東洋濾紙(株)製、２５ＡＳ０４５ＡＮ）で直ちにろ過した。
ろ過後の溶液に含まれるニッケル、コバルトの濃度は、誘導結合プラズマ発光分光分析装置（ＩＣＰ−ＡＥＳ，株式会社島津製作所製 ＩＣＰＳ−７５１０）を用いて定量した。検量線の作成には、関東化学株式会社の原子吸光分析用標準液（１０００mg/L）を適宜希釈して使用した。分析試料、及び検量線作成用標準試料は、全て硝酸酸性とした。ろ過後の溶液に含まれるセシウムの濃度は、イオンクロマトグラフ（ＤＩＯＮＥＸ製 ＤＸ−５００）を用いて定量した。分離カラムには、ＤＩＯＮＥＸ製「ＩｏｎＰａｃ(商標) ＣＧ１２Ａ」と「ＩｏｎＰａｃ(商標) ＣＳ１２Ａ」を使用し、溶離液には１８mmol/Lのメタンスルホン酸水溶液を使用した。溶離液の流速は１．０mL/minとした。
2.1 Measurement of adsorption amount Untreated basidiomycete, alkali-treated basidiomycete or ozone-treated basidiomycete was added at a rate of 1 g to 1 L of 1 ppm to 50 ppm metal solution. The mixed solution was stirred using a magnetic stirrer in a thermostat set at 30 ° C. The solution after the adsorption treatment was collected at any time and immediately filtered with a 0.45 μm acrylic polystyrene membrane filter (25AS045AN, manufactured by Toyo Roshi Kaisha, Ltd.).
The concentration of nickel and cobalt contained in the solution after filtration was quantified using an inductively coupled plasma optical emission spectrometer (ICP-AES, ICPS-7510 manufactured by Shimadzu Corporation). For the preparation of the calibration curve, a standard solution for atomic absorption analysis (1000 mg / L) from Kanto Chemical Co., Inc. was appropriately diluted. The analytical sample and the standard sample for preparing the calibration curve were all made acidic with nitric acid. The concentration of cesium contained in the solution after filtration was quantified using an ion chromatograph (DX-500 manufactured by DIONEX). “IonPac ™ CG12A” and “IonPac ™ CS12A” manufactured by DIONEX were used for the separation column, and 18 mmol / L methanesulfonic acid aqueous solution was used for the eluent. The flow rate of the eluent was 1.0 mL / min.
担子菌の乾燥重量当たりの金属イオン吸着量は、以下の（１）の式によって計算した。 The amount of metal ion adsorption per dry weight of basidiomycetes was calculated by the following equation (1).
式（１）：
Ｑ＝（Ｃｉ−Ｃｆ／ｍ）Ｖ
Formula (1):
Q = (Ci−Cf / m) V
ここで、Ｑは担子菌の乾燥重量（g）当たりの金属イオン吸着量（mg）、Ｃｉは金属イオンの初期濃度（mg/L）、Ｃｆは金属イオンの最終濃度（mg/L）、ｍは吸着に使用した担子菌の乾燥重量（g）、Ｖは反応混合物の体積（L）である。 Here, Q is the metal ion adsorption amount (mg) per dry weight (g) of basidiomycetes, Ci is the initial concentration of metal ions (mg / L), Cf is the final concentration of metal ions (mg / L), m Is the dry weight (g) of basidiomycetes used for adsorption, and V is the volume (L) of the reaction mixture.
結果を図１に示す。図１において各菌種（Pleurotus eryngii（エリンギ）、Hypsizygus marmoreus（ブナシメジ）、Lentinula edodes（シイタケ）、及びGrifola frondosa（マイタケ））のカラムのうち左が無処理担子菌、中央がアルカリ処理担子菌、右がオゾン処理担子菌の結果である。 The results are shown in FIG. In FIG. 1, among the columns of each bacterial species ( Pleurotus eryngii (Eringi), Hypsizygus marmoreus (Bunashimeji), Lentinula edodes (Shiitake), and Grifola frondosa (Maitake)), the left is an untreated basidiomycete, the center is an alkali-treated basidiomycete, The right is the result of ozone-treated basidiomycetes.
いずれの菌種でも、無処理担子菌で１．７〜２．８mg/gのニッケルを吸着しており、無処理でも高い吸着量を有することが示された。さらに、無処理担子菌とアルカリ処理担子菌のニッケル吸着量を比較するとPleurotus eryngiiではアルカリ処理７．２mg/g、無処理２．３mg/gであり、アルカリ処理担子菌のニッケル吸着量は無処理担子菌の３．１倍であった。同様に、Lentinula edodesではアルカリ処理４．８mg/gで無処理１．７mg/gの２．８倍、Hypsizygus marmoreusではアルカリ処理６．４mg/gで無処理２．８mg/gの２．３倍、及びGrifola frondosaではアルカリ処理４．９mg/gで無処理２．３mg/gの２．１倍であった。この結果から、本発明の吸着剤は、アルカリで処理する工程を行うことにより、ニッケル吸着量が大幅に増大することがわかった。特に、Pleurotus eryngiiはアルカリ処理によるニッケル吸着量の増大が顕著であり、無処理でのニッケル吸着量はHypsizygus marmoreusを下回っていたが、アルカリ処理をすることによってHypsizygus marmoreusの吸着量を０．８mg/g上回る結果を示した。 In any bacterial species, 1.7 to 2.8 mg / g of nickel was adsorbed by an untreated basidiomycete, and it was shown that a high adsorption amount was obtained even without treatment. Furthermore, when comparing the nickel adsorption amount of untreated basidiomycete and alkali-treated basidiomycete, Pleurotus eryngii is alkali-treated 7.2 mg / g and untreated 2.3 mg / g. It was 3.1 times that of basidiomycetes. Similarly, Lentinula edodes is 4.8 mg / g alkali-treated and 2.8 times the untreated 1.7 mg / g, Hypsizygus marmoreus is alkali-treated 6.4 mg / g and the untreated 2.8 mg / g 2.3 times. In Grifola frondosa , the alkali treatment was 4.9 mg / g, which was 2.1 times the non-treatment 2.3 mg / g. From this result, it was found that the adsorbent of the present invention greatly increases the amount of nickel adsorbed by performing the step of treatment with alkali. In particular, Pleurotus eryngii is remarkable increase in the nickel adsorption by alkaline treatment, nickel adsorption amount of untreated but were below Hypsizygus marmoreus, the adsorption amount of Hypsizygus marmoreus by the alkali treatment 0.8 mg / The result exceeded g.
糸状菌について、アルカリ溶液中で９５〜１００℃、４〜６時間の加熱処理を行い細胞壁のキチンを脱アセチル化させることで金属吸着量が増加することが報告されている（Muzzarelli R. A. A.ら：BiotechnoLBioeng, 22, 885-896 (1980)）。本発明の担子菌を含む吸着剤の製造において行ったアルカリ処理は室温で３０分間と温度が低く、時間も短いため、報告された糸状菌と同程度の脱アセチル化に至っていないと考えられるが、担子菌においても金属吸着量の増加にアルカリ処理が有効であることが示された。 It has been reported that filamentous fungi increase the amount of adsorbed metal by deacetylating cell wall chitin by heat treatment at 95-100 ° C. for 4-6 hours in an alkaline solution (Muzzarelli RAA et al .: BiotechnoLBioeng). , 22, 885-896 (1980)). Although the alkali treatment performed in the production of the adsorbent containing basidiomycetes of the present invention is as low as 30 minutes at room temperature and the time is short, it is considered that deacetylation has not been achieved to the same extent as the reported filamentous fungi. It was also shown that alkali treatment is effective for increasing the amount of metal adsorption in basidiomycetes.
アルカリ処理後にさらにオゾン処理を行った場合、ニッケル吸着量はそれぞれ、Pleurotus eryngiiで４．４mg/g、Lentinula edodesで２．６mg/g、Hypsizygus marmoreusで４．１mg/g及びGrifola frondosaで３．０mg/gであった。オゾン処理担子菌は、アルカリ処理担子菌と比較した場合、ニッケル吸着量の減少が見られたものの、いずれも無処理担子菌の１．３〜１．９倍の吸着量であった。したがって、オゾン処理でも一定の吸着量が期待できる。 When ozone treatment was further performed after alkali treatment, the amount of nickel adsorbed was 4.4 mg / g for Pleurotus eryngii , 2.6 mg / g for Lentinula edodes , 4.1 mg / g for Hypsizygus marmoreus , and 3.0 mg for Grifola frondosa , respectively. / g. The ozone-treated basidiomycetes showed a decrease in the amount of nickel adsorbed when compared to the alkali-treated basidiomycetes, but all were 1.3 to 1.9 times as much as the untreated basidiomycetes. Therefore, a constant amount of adsorption can be expected even with ozone treatment.
２．２ 吸着速度の測定
アルカリ処理担子菌（Pleurotus eryngii、Hypsizygus marmoreus、Lentinula edodes、Flammulina velutipes及びGrifola frondosa）を用いて、金属イオンを吸着する際の飽和吸着に達するまでの時間を調べた。使用した金属溶液は１０ppmのニッケル溶液（pH６）とした。サンプリングは吸着時間５分、１５分、３０分、４５分、６０分の５回実施し、それぞれ吸着後の溶液中の残存ニッケル濃度を上記と同様にして測定した。
2.2 Measurement of Adsorption Rate Using alkali-treated basidiomycetes ( Pleurotus eryngii , Hypsizygus marmoreus , Lentinula edodes , Flammulina velutipes and Grifola frondosa ), the time to reach saturation adsorption when metal ions were adsorbed was examined. The metal solution used was a 10 ppm nickel solution (pH 6). Sampling was carried out 5 times, 15 minutes, 30 minutes, 45 minutes and 60 minutes for the adsorption time, and the residual nickel concentration in the solution after adsorption was measured in the same manner as described above.
結果を図２に示す。ニッケルは、いずれの担子菌においても吸着開始５分後までに概ね吸着平衡に達しており、水溶液中のニッケルイオンは担子菌に速やかに吸着されることが明らかになった。また、菌種によって金属イオンの吸着量は異なるが、吸着に要する時間には大きな差がないことがわかった。 The results are shown in FIG. Nickel almost reached the adsorption equilibrium by 5 minutes after the start of adsorption in any basidiomycete, and it became clear that nickel ions in the aqueous solution were rapidly adsorbed by the basidiomycete. Moreover, although the amount of metal ion adsorption differs depending on the bacterial species, it was found that there was no significant difference in the time required for adsorption.
２．３ 吸着特性
アルカリ処理担子菌による金属吸着特性をさらに調べた。吸着剤としてアルカリ処理担子菌（Pleurotus eryngii、Hypsizygus marmoreus、Lentinula edodes、Flammulina velutipes及びGrifola frondosa）を用い、吸着質としてニッケル、コバルト、リチウム、ホウ素、セシウムを含む金属溶液を用いた。金属濃度はそれぞれ１ppm、５ppm、１０ppm、２５ppm、５０ppmとした。pH６の金属溶液１Lに対してアルカリ処理担子菌を１gの割合で投入し、混合液とした。混合液は３０℃に設定した恒温器内においてマグネチックスターラーで攪拌した。ニッケル溶液を用いた吸着速度の実験結果から、すべての担子菌菌種で吸着平衡に達する時間よりも長い３０分間を攪拌時間として設定した。
2.3 Adsorption characteristics The metal adsorption characteristics of the alkali-treated basidiomycetes were further investigated. An alkali-treated basidiomycete ( Pleurotus eryngii , Hypsizygus marmoreus , Lentinula edodes , Flammulina velutipes and Grifola frondosa ) was used as the adsorbent, and a metal solution containing nickel, cobalt, lithium, boron and cesium was used as the adsorbate. The metal concentrations were 1 ppm, 5 ppm, 10 ppm, 25 ppm and 50 ppm, respectively. An alkali-treated basidiomycete was added at a rate of 1 g to 1 L of a pH 6 metal solution to prepare a mixed solution. The liquid mixture was stirred with a magnetic stirrer in a thermostat set to 30 ° C. From the experimental results of the adsorption rate using the nickel solution, 30 minutes longer than the time to reach the adsorption equilibrium in all basidiomycete species was set as the stirring time.
各種担子菌の金属吸着等温線を図３〜５に示す。
また、Langmuir式及びFreundlich式の２種類の吸着等温式を用いて吸着等温線の解析を行った。Langmuirモデルは式（２）に従った。
The metal adsorption isotherms of various basidiomycetes are shown in FIGS.
In addition, the adsorption isotherm was analyzed using two types of adsorption isotherms, the Langmuir equation and the Freundlich equation. The Langmuir model followed equation (2).
式（２）：
Ｗ＝ａＷS Ｃ／（１＋ａＣ）
Formula (2):
W = aW S C / (1 + aC)
ここで、Ｗは担子菌単位重量当たりの吸着量（mg/g）、Ｃは溶質の平衡濃度（mg/L）、ＷSを飽和吸着量、ａは吸着平衡定数とする。
Langmuir式（２）を変形した直線式（３）に測定値をあてはめてＣ及びＣ／Ｗでプロットし、直線関係が成立している場合には直線の傾きと切片から吸着平衡定数及び飽和吸着量を算出した。
Here, W is the amount of adsorption per unit weight of basidiomycetes (mg / g), C is the equilibrium concentration of solute (mg / L), W S is the saturated adsorption amount, and a is the adsorption equilibrium constant.
The measured value is applied to the linear equation (3) modified from the Langmuir equation (2) and plotted in C and C / W. If the linear relationship is established, the adsorption equilibrium constant and saturated adsorption are determined from the slope and intercept of the straight line. The amount was calculated.
式（３）：
Ｃ／Ｗ＝（１／ａＷS）＋（１／ＷS）Ｃ
Formula (3):
C / W = (1 / aW S ) + (1 / W S ) C
Freundlichモデルは式（４）にしたがった。 The Freundlich model was according to equation (4).
式（４）：
Ｗ＝ＫＣ1/n
Formula (4):
W = KC 1 / n
ここで、Ｋ及び１／ｎは吸着定数である。両辺の対数をとると式（５）のような直線一次式が得られる。その直線の傾きから１／ｎが得られ、Ｃ＝１のときの吸着量からＫを求めた。 Here, K and 1 / n are adsorption constants. Taking the logarithm of both sides yields a linear linear expression such as the expression (5). 1 / n was obtained from the slope of the straight line, and K was determined from the amount of adsorption when C = 1.
式（５）：
log Ｗ＝log Ｋ＋（１／ｎ）log Ｃ
Formula (5):
log W = log K + (1 / n) log C
前処理をした担子菌５菌種での重金属ニッケル、コバルト、セシウムの吸着等温線をLangmuir式及びFreundlich式によって解析した吸着パラメーターを表１に示した。 Table 1 shows the adsorption parameters obtained by analyzing the adsorption isotherms of heavy metal nickel, cobalt, and cesium by the pretreated basidiomycetes using the Langmuir equation and the Freundlich equation.
Langmuir式の吸着等温線で解析した結果、ニッケルの飽和吸着量はPleurotus eryngii＞Hypsizygus marmoreus＞Lentinula edodes＞Flammulina velutipes＞Grifola frondosという結果を示し、Pleurotus eryngiiとHypsizygus marmoreusの飽和吸着量は８．０mg/gを上回った。コバルトではPleurotus eryngii＞Hypsizygus marmoreus＞Lentinula edodes＞Grifola frondosa＞Flammulina velutipesという結果を示し、最も吸着量が多いPleurotus eryngiiで７．９６mg/g、次いでHypsizygus marmoreusで６．８６mg/gであった。セシウムでは、Grifola frondos＞Hypsizygus marmoreus＞Lentinula edodes＞Pleurotus eryngii＞Flammulina velutipesという結果を示した。Hypsizygus marmoreusはニッケル、コバルト、セシウムのいずれも比較的高い飽和吸着量を示した。一方、Pleurotus eryngii及びGrifola frondosaでは重金属を選択的に吸着する傾向が見られた。すなわち、Pleurotus eryngiiではニッケル及びコバルトで高い吸着能が見られたが、セシウムでは飽和吸着量が低く、反対にGrifola frondosaではセシウムの吸着能が高く、ニッケル及びコバルトの飽和吸着量が比較的低くなる結果であった。 As a result of analysis using the Langmuir-type adsorption isotherm, the saturated adsorption amount of nickel was Pleurotus eryngii > Hypsizygus marmoreus > Lentinula edodes > Flammulina velutipes > Grifola frondos , and the saturated adsorption amount of Pleurotus eryngii and Hypsizygus marmoreus was 8.0 mg / hypuszygus marmoreus. exceeded g. Cobalt showed the following results: Pleurotus eryngii > Hypsizygus marmoreus > Lentinula edodes > Grifola frondosa > Flammulina velutipes , with Pleurotus eryngii with the highest amount of adsorption, 7.96 mg / g and then Hypsizygus marmoreus with 6.86 mg / g. For cesium, Grifola frondos > Hypsizygus marmoreus > Lentinula edodes > Pleurotus eryngii > Flammulina velutipes . Hypsizygus marmoreus showed relatively high saturated adsorption for all of nickel, cobalt and cesium. On the other hand, Pleurotus eryngii and Grifola frondosa showed a tendency to selectively adsorb heavy metals. In other words, Pleurotus eryngii showed high adsorption capacity with nickel and cobalt, while cesium had low saturated adsorption capacity, while Grifola frondosa had high cesium adsorption capacity and relatively low nickel and cobalt saturated adsorption capacity. It was a result.
このことから、担子菌の菌種を選択することで廃水中から目的の重金属を高濃度に回収できる可能性があることが推察された。また、Langmuir式の吸着等温線解析ではいずれの担子菌及び重金属の組み合わせにおいても０．９以上の高い相関性が得られた。Langmuir式は単分子層での吸着及び化学吸着の挙動を示す式であることから、担子菌への重金属の吸着メカニズムは化学吸着であることが明らかになった。 From this, it was speculated that the target heavy metal may be recovered in high concentration from the wastewater by selecting the basidiomycete species. Moreover, in the Langmuir-type adsorption isotherm analysis, a high correlation of 0.9 or higher was obtained for any combination of basidiomycetes and heavy metals. The Langmuir equation is an equation showing the behavior of adsorption and chemisorption in monolayers, and it was clarified that the adsorption mechanism of heavy metals to basidiomycete is chemisorption.
Freundlich式によって計算した吸着パラメーターＫ値は大きいほど吸着能が大きくなり、ｎ値は吸着強度を表している。ニッケル、コバルトは吸着特性が類似しており、いずれもPleurotus eryngiiが最も吸着能が高く、次いでHypsizygus marmoreus、Lentinula edodesと続くことがわかった。ｎ値の比較からLentinula edodes、Grifola frondosの吸着強度が比較的高いことが示された。このことから、担子菌に吸着した重金属を回収する際、親和力が小さい３菌種、Pleurotus eryngii、Hypsizygus marmoreus、Flammulina velutipesは回収効率が比較的良いことが推察された。 The larger the adsorption parameter K value calculated by the Freundlich equation, the larger the adsorption ability, and the n value represents the adsorption strength. Nickel and cobalt have similar adsorption characteristics, and Pleurotus eryngii has the highest adsorption capacity, followed by Hypsizygus marmoreus and Lentinula edodes . Comparison of n values indicated that the adsorption strength of Lentinula edodes and Grifola frondos was relatively high. This suggests that when recovering heavy metals adsorbed on basidiomycetes, three species with low affinity, Pleurotus eryngii , Hypsizygus marmoreus and Flammulina velutipes , have relatively good recovery efficiency.
なお、リチウム、ホウ素についても、５種類のアルカリ処理担子菌を用いて同様の実験を行ったが、いずれの担子菌に対しても吸着は全く見られなかった。リチウムイオンは水素イオンやナトリウムイオンよりも陽イオン交換樹脂への吸着性が低いことが知られており、担子菌における吸着メカニズムがイオン交換樹脂等と同様の反応による可能性が考えられる。一方、ホウ素は水溶液中で１価の陰イオンとなるが、担子菌体は陰イオンを吸着できる官能基を保有していないものと推察される。 For lithium and boron, the same experiment was performed using five types of alkali-treated basidiomycetes, but no adsorption was observed for any basidiomycete. Lithium ions are known to be less adsorbable to cation exchange resins than hydrogen ions and sodium ions, and the adsorption mechanism in basidiomycetes may be due to the same reaction as ion exchange resins. On the other hand, boron becomes a monovalent anion in an aqueous solution, but it is assumed that basidiomycetes do not have a functional group capable of adsorbing anions.
３．吸着した重金属の脱離回収
前記１．と同様にして、４菌種の担子菌（Pleurotus eryngii、Hypsizygus marmoreus、Grifola frondosa、Flammulina velutipes）を用いてアルカリ処理担子菌を製造し、前記２．１と同様にしてニッケルを吸着させ、吸着量を測定した。Pleurotus eryngii、Hypsizygus marmoreus、Grifola frondosa、Flammulina velutipesのそれぞれのニッケル吸着量は５．６３、５．４５、４．０８及び３．８４mg/gであった。
その後、アルカリ処理担子菌に吸着させたニッケルの脱離回収を、酸性溶液を用いて行った。酸性溶液としては、塩酸（関東化学(株)製、有害金属測定用、濃度３５〜３７％）及び蒸留水を使用してｐＨ１、２、３、及び４に調整したものを用いた。重金属の脱離法は、酸性溶液１Lに対して、重金属吸着後の担子菌含有廃菌床１gを投入し、室温、３０分間マグネチックスターラーで攪拌して行った。溶液中のニッケルイオン濃度はＩＣＰ-ＡＥＳを用いて定量した。回収率を、以下の計算式によって算出した：
回収率（％）＝回収量／吸着量×１００
3. Desorption recovery of adsorbed heavy metals In the same manner as above , alkali-treated basidiomycetes were produced using basidiomycetes of 4 bacterial species ( Pleurotus eryngii , Hypsizygus marmoreus , Grifola frondosa , Flammulina velutipes ), and nickel was adsorbed in the same manner as in 2.1 above. Was measured. The nickel adsorption amounts of Pleurotus eryngii , Hypsizygus marmoreus , Grifola frondosa and Flammulina velutipes were 5.63 , 5.45 , 4.08 and 3.84 mg / g, respectively.
Thereafter, desorption and recovery of nickel adsorbed on the alkali-treated basidiomycete was performed using an acidic solution. As the acidic solution, those adjusted to pH 1, 2, 3, and 4 using hydrochloric acid (manufactured by Kanto Chemical Co., Inc., for measuring harmful metals, concentration 35 to 37%) and distilled water were used. The heavy metal desorption method was performed by adding 1 g of basidiomycetous-containing waste fungus bed after adsorption of heavy metal to 1 L of acidic solution and stirring with a magnetic stirrer at room temperature for 30 minutes. The nickel ion concentration in the solution was quantified using ICP-AES. The recovery rate was calculated by the following formula:
Recovery rate (%) = recovered amount / adsorbed amount × 100
酸性溶液のｐＨ条件の違いによるニッケル回収率を菌種ごとに調べた結果を、図６に示す。酸性溶液がｐＨ１のときの回収率は、それぞれ、Pleurotus eryngiiで９８．２％、Hypsizygus marmoreusで９７．６％、Lentinula edodesで９０．７％、Flammulina velutipesで９６．４％であった。ｐＨ３以下の酸性溶液におけるニッケル回収率の変動はあまり大きくなく、ｐＨ３以下の酸性溶液を用いることで、いずれの担子菌においても吸着したニッケルを約９０％以上の高い効率で回収できることが明らかになった。 FIG. 6 shows the results of examining the nickel recovery rate for each bacterial species according to the difference in the pH conditions of the acidic solution. The recoveries when the acidic solution was pH 1 were 98.2% for Pleurotus eryngii , 97.6% for Hypsizygus marmoreus , 90.7% for Lentinula edodes , and 96.4% for Flammulina velutipes , respectively. The fluctuation of the nickel recovery rate in the acidic solution at pH 3 or less is not so large, and it becomes clear that the nickel adsorbed in any basidiomycete can be recovered with high efficiency of about 90% or more by using the acidic solution at pH 3 or less. It was.
一方、pHが４に上がるといずれの菌種でもニッケル回収率が急激に下がった。これは、担子菌の陽イオン吸着サイトに対して重金属イオンと水素イオンが競合し、pH３付近ではプロトン濃度の上昇に伴い陽イオン吸着サイトの多くがプロトン化し、重金属イオンの脱離が進むことに起因すると考えられる。 On the other hand, when the pH increased to 4, the nickel recovery rate rapidly decreased in any bacterial species. This is because heavy metal ions and hydrogen ions compete with the cation adsorption sites of basidiomycetes, and as the proton concentration increases, most of the cation adsorption sites are protonated and the desorption of heavy metal ions proceeds. It is thought to be caused.
また、Freundlichモデルでの吸着等温線解析から得られたニッケル吸着強度と回収率との関係を図７に示した。ニッケルの吸着強度と回収率との間にはほぼ負の相関関係が得られ、ニッケルの吸着強度が最も高いLentinula edodesのニッケル回数率が最も低い結果となった。これにより、吸着強度が重金属回収率に影響を及ぼす可能性が示唆された。 FIG. 7 shows the relationship between the nickel adsorption strength and the recovery rate obtained from the adsorption isotherm analysis using the Freundlich model. There was a nearly negative correlation between the adsorption strength and the recovery rate of nickel, and the nickel frequency rate of Lentinula edodes with the highest nickel adsorption strength was the lowest. This suggested that the adsorption strength may affect the heavy metal recovery rate.
４．細孔分布測定及び吸着メカニズムの解明
水銀圧入式細孔分布測定装置（島津製作所製、オートポアIV９５００）を用いて、子実体の無処理担子菌、アルカリ処理担子菌（いずれもFlammulina velutipes）の細孔分布図をそれぞれ求めた。水銀と試料の接触角を１４０度として細孔分布を算出した。
4). Measurement of pore distribution and elucidation of adsorption mechanism Using mercury intrusion type pore distribution measuring device (manufactured by Shimadzu Corp., Autopore IV9500), the pores of untreated basidiomycetes of fruit bodies and alkali-treated basidiomycetes (both Flammulina velutipes ) Each distribution map was obtained. The pore distribution was calculated with the contact angle of mercury and the sample being 140 degrees.
水銀圧入式細孔分布測定法による無処理担子菌及びアルカリ処理担子菌の細孔分布を図８に示す。無処理担子菌（図８−Ａ）では複数見られた細孔のピークが、アルカリ処理担子菌（図８−Ｂ）においては１５μm付近に集中することがわかった。重金属吸着量はアルカリ処理によって増加したが、担子菌へのアルカリ処理前後で物理吸着に必要な微細孔の発達は見られなかった。
なお、アルカリ処理担子菌（Flammulina velutipes）の走査型電子顕微鏡像（ＳＥＭ、エリオニクス社製、ＥＲＡ−８９００ＦＥ、２５０倍）を図９に示した。
FIG. 8 shows the pore distribution of untreated basidiomycete and alkali-treated basidiomycete by mercury intrusion pore distribution measurement method. It was found that a plurality of pore peaks observed in the untreated basidiomycete (FIG. 8-A) are concentrated around 15 μm in the alkali-treated basidiomycete (FIG. 8-B). Although the amount of heavy metal adsorption increased with alkali treatment, the development of micropores necessary for physical adsorption was not observed before and after alkali treatment on basidiomycetes.
A scanning electron microscope image (SEM, manufactured by Elionix, ERA-8900FE, 250 times) of alkali-treated basidiomycete ( Flammmulina velutipes ) is shown in FIG.
担子菌の細胞壁は主にグルカン、キチン、タンパク質からなり、菌体表面にアミノ基及び水酸基等の陽イオンを吸着できる官能基を有している。一方、重金属イオンは水溶液中で電荷をもっており、実験の結果から、担子菌に吸着した重金属がｐＨ４以下、特にｐＨ３以下の酸性溶液中で効率的に脱離することがわかった。また、吸着等温線解析の結果から、化学吸着で用いるLangmuir式の吸着等温式が成り立つことがわかった。
これらのことから、担子菌が重金属イオンを吸着する主なメカニズムはイオン間相互作用による化学吸着であることが示された。
The basidiomycete cell wall is mainly composed of glucan, chitin, and protein, and has functional groups capable of adsorbing cations such as amino groups and hydroxyl groups on the surface of the cells. On the other hand, heavy metal ions have a charge in an aqueous solution, and the results of the experiment show that heavy metals adsorbed on basidiomycetes are efficiently desorbed in an acidic solution having a pH of 4 or less, particularly pH 3 or less. In addition, from the results of adsorption isotherm analysis, it was found that the Langmuir-type adsorption isotherm used in chemical adsorption holds.
From these, it was shown that the main mechanism by which basidiomycetes adsorb heavy metal ions is chemisorption by the interaction between ions.
５．吸着剤の製造方法の検討
Pleurotus eryngii（エリンギ）及びLentinula edodes（シイタケ）の担子菌類２菌種の子実体を用い、前記１．に記載したとおりに、無処理担子菌、アルカリ処理担子菌を製造した。
5. Examination of manufacturing method of adsorbent
Using fruit bodies of two basidiomycetes of Pleurotus eryngii (Eringi) and Lentinula edodes (Shiitake), As described in 1., untreated basidiomycetes and alkali-treated basidiomycetes were produced.
すなわち、まず、子実体を、乾燥機で６０℃、３日間乾燥させ、それを粉砕機で２０秒間粉砕し、目開き３５５μmのふるい通過分をさらに６０℃、２４時間乾燥させた（「無処理担子菌」）。
無処理担子菌１０gを０．２M 水酸化ナトリウム溶液１Lに投入し、室温で３０分間マグネチックスターラーを用いて撹拌することにより、アルカリ処理を行った。この菌体を、洗浄液のｐＨが中性域に達するまでイオン交換水を用いて洗浄した。その後、遠心分離及び吸引ろ過によって菌体を回収し、乾燥器で６０℃、２日間乾燥させ、それを粉砕機で粉砕し、目開き３５５μmのふるい通過分をさらに６０℃、２４時間乾燥させた（「アルカリ処理担子菌」）。
That is, first, the fruiting bodies were dried at 60 ° C. for 3 days with a dryer, pulverized with a pulverizer for 20 seconds, and the sieve passage having a mesh size of 355 μm was further dried at 60 ° C. for 24 hours (“No treatment” Basidiomycetes ").
10 g of untreated basidiomycetes were put into 1 L of 0.2M sodium hydroxide solution and stirred with a magnetic stirrer at room temperature for 30 minutes for alkali treatment. The cells were washed with ion exchange water until the pH of the washing solution reached a neutral range. Thereafter, the cells were collected by centrifugation and suction filtration, dried at 60 ° C. for 2 days with a dryer, pulverized with a pulverizer, and further dried through a sieve having an opening of 355 μm at 60 ° C. for 24 hours. ("Alkaline treated basidiomycetes").
アルカリ処理担子菌の一部について、さらにオゾン処理を行った。１０Lのガラス製デシケータ内に担子菌０．１gを静置し、オゾン発生装置（日本オゾン株式会社製、ＯＮ−３−２型）を用いて発生させたオゾンをデシケータ内に導入した。オゾンの発生量は３．０mg/minでオゾンの導入は約１時間とした（「アルカリ処理後オゾン処理担子菌」とする。前記１．における「オゾン処理担子菌」に相当する）。 Ozone treatment was further performed on some of the alkali-treated basidiomycetes. 0.1 g of basidiomycetes were allowed to stand in a 10 L glass desiccator, and ozone generated using an ozone generator (manufactured by Nippon Ozone Co., Ltd., ON-3-2 type) was introduced into the desiccator. The amount of ozone generated was 3.0 mg / min, and ozone was introduced for about 1 hour (referred to as “ozone-treated basidiomycete after alkali treatment”, which corresponds to “ozone-treated basidiomycete” in 1.).
アルカリ処理及びオゾン処理の順序を変えて吸着剤を製造した。すなわち、無処理担子菌１０ｇを、１０Lのガラス製デシケータ内に静置し、オゾン発生装置（日本オゾン株式会社製、ＯＮ−３−２型）を用いて発生させたオゾンをデシケータ内に導入した。オゾンの発生量は３．０mg/minでオゾンの導入は約１時間とした。その後、オゾン処理した無処理担子菌１０gを０．２M 水酸化ナトリウム溶液１Lに投入し、室温で３０分間マグネチックスターラーを用いて撹拌することにより、アルカリ処理を行った。この菌体を、洗浄液のｐＨが中性域に達するまでイオン交換水を用いて洗浄した。その後、遠心分離及び吸引ろ過によって菌体を回収し、乾燥器で６０℃、２日間乾燥させ、それを粉砕機で粉砕し、目開き３５５μmのふるい通過分をさらに６０℃、２４時間乾燥させた（「オゾン処理後アルカリ処理担子菌」とする）。 Adsorbents were produced by changing the order of alkali treatment and ozone treatment. That is, 10 g of untreated basidiomycetes were allowed to stand in a 10 L glass desiccator, and ozone generated using an ozone generator (made by Nippon Ozone Co., Ltd., ON-3-2 type) was introduced into the desiccator. . The amount of ozone generated was 3.0 mg / min, and ozone was introduced for about 1 hour. Thereafter, 10 g of untreated basidiomycete treated with ozone was added to 1 L of 0.2 M sodium hydroxide solution, and stirred with a magnetic stirrer for 30 minutes at room temperature to perform alkali treatment. The cells were washed with ion exchange water until the pH of the washing solution reached a neutral range. Thereafter, the cells were collected by centrifugation and suction filtration, dried at 60 ° C. for 2 days with a dryer, pulverized with a pulverizer, and further dried through a sieve having an opening of 355 μm at 60 ° C. for 24 hours. ("Alkaline treated basidiomycete after ozone treatment").
前記２．1と同様にして、各吸着剤を用いて１０ppmのニッケル（Ｎｉ2+）を含む金属溶液からの金属の吸着量を測定した（吸着処理のための攪拌時間は３０分間とした）。結果を表２に示す。 In the same manner as in 2.1 above, the amount of metal adsorbed from a metal solution containing 10 ppm of nickel (Ni 2+ ) was measured using each adsorbent (the stirring time for the adsorption treatment was 30 minutes). . The results are shown in Table 2.
オゾン処理後にアルカリ処理をした場合は、アルカリ処理後にオゾン処理をしたものと比較して、ニッケル吸着量がシイタケで２０８％、エリンギで１６８％となることがわかった。したがって、アルカリ及びオゾンによる処理を行う順番が吸着量に大きな影響を与えること、及びオゾン処理後にアルカリ処理を行うことによって各担子菌による金属吸着量が顕著に増大することが明らかになった。 When the alkali treatment was performed after the ozone treatment, it was found that the nickel adsorption amount was 208% for shiitake and 168% for eringi, compared with the case where the ozone treatment was performed after the alkali treatment. Therefore, it has been clarified that the order of treatment with alkali and ozone has a great influence on the amount of adsorption, and that the amount of metal adsorption by each basidiomycete is significantly increased by performing the alkali treatment after the ozone treatment.
６．担子菌を含む廃菌床を用いた吸着剤の製造及び金属吸着量の測定
Lentinula edodes（シイタケ）を含む、シイタケ栽培後の廃菌床を用いたこと以外は前記５．と同様にして、「無処理担子菌」、「アルカリ処理担子菌」及び「オゾン処理後アルカリ処理担子菌」を製造した。この各種吸着剤を用いて、前記５．と同様にしてニッケル吸着量を測定し、子実体を用いて製造した吸着剤による吸着量と比較した。結果を表３に示す。
6). Production of adsorbent using waste bed containing basidiomycetes and measurement of metal adsorption
4. The above except that the waste fungus bed after shiitake cultivation including Lentinula edodes (shiitake) was used. In the same manner, “untreated basidiomycetes”, “alkali-treated basidiomycetes”, and “ozone-treated alkali-treated basidiomycetes” were produced. Using these various adsorbents, 5. The amount of nickel adsorbed was measured in the same manner as described above and compared with the amount adsorbed by the adsorbent produced using the fruit body. The results are shown in Table 3.
シイタケを栽培した後の廃菌床を用いた吸着剤は、シイタケの子実体を用いた吸着剤よりも多量のニッケルを吸着することが明らかになった。また、廃菌床を用いた吸着剤の場合も、子実体を用いた吸着剤と同様に、無処理のものと比較してアルカリ処理を施した場合の方が吸着量が多く、オゾン処理後にアルカリ処理を施すことによってさらに吸着量が増大することがわかった。これは、オゾン処理で酸化分解することにより吸着剤表面の陽イオン吸着サイトが増加し、吸着量が増加したためである可能性が考えられる。 It was revealed that the adsorbent using the waste fungus bed after cultivation of shiitake adsorbs more nickel than the adsorbent using the fruit body of shiitake. Also, in the case of the adsorbent using the waste fungus bed, the amount of adsorption is higher when the alkali treatment is applied compared to the untreated one, after the ozone treatment. It was found that the amount of adsorption increased further by applying alkali treatment. This may be because the cation adsorption sites on the adsorbent surface increased due to oxidative decomposition by ozone treatment, and the adsorption amount increased.
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