JP2018150261A - Antibacterial/antifungal component derived from soil and separation method thereof - Google Patents
Antibacterial/antifungal component derived from soil and separation method thereof Download PDFInfo
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本発明は、新規な土壌由来の抗菌活性を有する無機金属複合体に関する。本発明はまた、そのような抗菌性無機金属複合体を有する抗菌剤に関する。 The present invention relates to a novel soil-derived inorganic metal composite having antibacterial activity. The present invention also relates to an antibacterial agent having such an antibacterial inorganic metal composite.
リモナイトとは、沼地や浅い海などの鉄分を多く含む水が空気に触れ、沈殿作用がおきて沈殿した黄土のことである。リモナイトはまた、「褐鉄鉱」、「沼鉄鉱」とも呼ばれ、酸化鉱物系のレピドクロサイトやゲーサイトなどが混ざった鉱物の総称である。リモナイトは、暗褐色または黒色の団塊、土状のものとして産出され、その化学組成はＦｅＯ（ＯＨ）・ｎＨ2Ｏであり、そのため、褐鉄鋼としても用いられている。 Limonite is a loess that has settled due to the precipitation of water that contains a lot of iron, such as marshes and shallow seas. Limonite is also called “limonite” and “numanite”, and is a general term for minerals containing mixed oxide minerals such as lipid dodecite and goethite. Limonite is produced in the form of dark brown or black nodules and soils, and its chemical composition is FeO (OH) .nH 2 O, and is therefore also used as brown iron steel.
阿蘇・狩尾一帯に分布している「阿蘇の黄土」を採掘後、３年間熟成させた土壌であるリモナイト（褐鉄鉱）は、硫化水素吸着剤、農業用土壌活性土、畜産用飼料、ペット用健康消臭補助製品、水質活性土などに利用されている。また、昔より、牛飼いから牛の下痢止めとして重宝されてきた。阿蘇の黄土は鉄分含有量が多く、その他、カルシウム、マグネシウムといったマルチミネラルがバランスよく含まれている。 Limonite (Limonite), a soil that has been aged for 3 years after mining “Aso Loess” distributed in the Aso / Kao region, is composed of hydrogen sulfide adsorbent, agricultural soil activated soil, livestock feed, and pet use. It is used in health deodorant auxiliary products and water quality activated soil. In addition, it has been useful for a long time as a cow diarrhea prevention from the cow owner. Aso's ocher has a high iron content, and it also contains a good balance of multi minerals such as calcium and magnesium.
従来より、銀、銅、亜鉛等の無機金属塩類及び有機金属塩類が抗菌・殺菌性に優れていることが知られており、銀、銅、亜鉛等の抗菌性金属のうち特に銀イオンを含む無機金属塩類は、安全性が比較的高く顕著な抗菌力を示すことから無機系抗菌剤として注目を集めている。 Conventionally, it has been known that inorganic metal salts and organic metal salts such as silver, copper and zinc are excellent in antibacterial and bactericidal properties, and silver ions are included among antibacterial metals such as silver, copper and zinc. Inorganic metal salts are attracting attention as inorganic antibacterial agents because they are relatively safe and exhibit remarkable antibacterial activity.
またこれまでに、土壌中の抗菌や抗真菌成分として、明礬類や腐植泥製品／ヒューマス・カルミーが知られており、ヒューマス・カルミーより分離された抗菌性無機金属複合体としてＫＨ−Ｃ1が知られている（非特許文献１：日本農芸化学会誌, ６４巻、第４号、９０７頁〜９１２頁、１９９０年）。
ＫＨ−Ｃ1は、無色針状晶で、水、メタノールおよびエタノールには可溶であるが、アセトン、ヘキサンおよびエーテルなので有機溶媒には不溶である。ＫＨ−Ｃ1は、Ｆｅ，Ｎａ，ＡｌおよびＭｇを含み、ＣおよびＮを全く含まない無機塩の硫酸塩からなる複合体である。また、ＫＨ−Ｃ1は、明礬類には含まれていないマグネシウムや２価の鉄を含み、カリウムや窒素を全く含まず、カリウム、アンモニウム、鉄（３価）およびナトリウムの明礬類とは金属組成比も異なる。
So far, alum and humus mud products / Hummas Karmy have been known as antibacterial and antifungal ingredients in soil, and KH-C 1 is an antibacterial inorganic metal complex separated from Hughs Karmy. (Non-patent document 1: Journal of Japanese Society for Agricultural Chemistry, 64, No. 4, pages 907-912, 1990).
KH-C 1 is a colorless needle crystal and is soluble in water, methanol and ethanol, but is insoluble in organic solvents since it is acetone, hexane and ether. KH-C 1 is a composite composed of a sulfate of an inorganic salt containing Fe, Na, Al, and Mg and not containing C and N at all. KH-C 1 contains magnesium and divalent iron, which are not contained in alum, and does not contain potassium or nitrogen at all, and potassium, ammonium, iron (trivalent) and sodium alum are metals The composition ratio is also different.
本発明は新規な抗菌性無機金属複合体を提供することである。より具体的には、リモナイトを利用して、優れた抗菌・抗真菌作用を有する新規な無機金属複合体を提供することである。 The present invention is to provide a novel antibacterial inorganic metal composite. More specifically, it is to provide a novel inorganic metal composite having an excellent antibacterial / antifungal action using limonite.
本発明者らは、リモナイトに抗菌作用があることを見出し、そしてさらに、リモナイトから優れた抗菌活性をもつ特定の無機金属の複合体を分離することに成功し、本発明を完成させた。
すなわち、本発明は以下のものを提供する。
（１）抗菌および抗真菌活性を有する、下記：
（ａ）主要な金属として、Ａｌ，Ｍｎ，Ｚｎ，およびＣａを含む、
（ｂ）前記金属の含有量は、Ａｌ＞Ｍｎ＞Ｚｎ，Ｃａである、および
（ｃ）グラム陽性菌、グラム陰性菌および真菌に対して抗菌活性をもつ、
の特徴を有する抗菌性無機金属複合体。
（２）前記無機金属複合体は、以下の式：
［Ａｌ，Ｍｎ，Ｃａ，ＳＯ4，Ｆ］・ｎＨ2Ｏ
であらわされる、硫化塩およびフッ化物を含む複塩である、上記（１）に記載の抗菌性無機金属複合体。
（３）リモナイトから抽出された上記（１）または（２）に記載の抗菌性無機金属複合体。
（４）前記抗菌活性が、グラム陽性菌、グラム陰性菌および真菌のいずれに対しても、ＭＩＣ値で２ｍｇ／ｍｌ以下（好ましくは１ｍｇ／ｍｌ以下）である上記（１）〜（３）のいずれか一つに記載の抗菌性無機金属複合体。
（５）結晶形として板状結晶を形成する上記（１）〜（４）のいずれか一つに記載の抗菌性無機金属複合体。
The present inventors have found that limonite has an antibacterial action, and further succeeded in separating a specific inorganic metal complex having excellent antibacterial activity from limonite, thereby completing the present invention.
That is, the present invention provides the following.
(1) Having antibacterial and antifungal activity, the following:
(A) including Al, Mn, Zn, and Ca as main metals,
(B) the content of the metal is Al>Mn> Zn, Ca, and (c) has antibacterial activity against gram positive bacteria, gram negative bacteria and fungi,
An antibacterial inorganic metal composite having the following characteristics:
(2) The inorganic metal composite has the following formula:
[Al, Mn, Ca, SO 4 , F] · nH 2 O
The antibacterial inorganic metal composite according to (1) above, which is a double salt containing a sulfide and a fluoride.
(3) The antibacterial inorganic metal composite according to (1) or (2) extracted from limonite.
(4) The antibacterial activity according to (1) to (3) above, wherein the antibacterial activity is 2 mg / ml or less (preferably 1 mg / ml or less) in terms of MIC for any of Gram positive bacteria, Gram negative bacteria and fungi The antibacterial inorganic metal composite according to any one of the above.
(5) The antibacterial inorganic metal composite according to any one of (1) to (4), wherein a plate-like crystal is formed as a crystal form.
（６）リモナイトから以下の工程：
（ｉ）リモナイトを水に懸濁し、水抽出物を回収する工程、
（ｉｉ）前記水抽出物から、有機溶媒可溶物を除く工程、および
（ｉｉｉ）上記工程（ｉｉ）により得られた水抽出物からメタノール可溶物を除く工程、
により抽出される抗菌性無機金属複合体。
（７）前記有機溶媒可溶物が、アセトン可溶物、エーテル可溶物およびクロロホルム可溶物である、上記（６）に記載の抗菌性無機金属複合体。
（８）前記リモナイトが、リモナイトＬＭＢ３００である、上記（６）または（７）に記載の抗菌性無機金属複合体。
（９）前記抗菌性無機金属複合体が板状結晶として回収される、上記（６）〜（８）のいずれか一つに記載の抗菌性無機金属複合体。
(6) The following steps from limonite:
(I) suspending limonite in water and recovering the water extract;
(Ii) a step of removing an organic solvent soluble material from the water extract, and (iii) a step of removing a methanol soluble material from the water extract obtained by the step (ii),
Antibacterial inorganic metal composites extracted by
(7) The antibacterial inorganic metal composite according to (6), wherein the organic solvent soluble material is an acetone soluble material, an ether soluble material, or a chloroform soluble material.
(8) The antibacterial inorganic metal composite according to (6) or (7), wherein the limonite is limonite LMB300.
(9) The antibacterial inorganic metal composite according to any one of (6) to (8), wherein the antibacterial inorganic metal composite is recovered as a plate crystal.
（１０）以下の工程、
（ｉ）リモナイトを水に懸濁し、水抽出物を回収する工程、
（ｉｉ）前記水抽出物から、有機溶媒可溶物を除く工程、
（ｉｉｉ）上記工程（ｉｉ）により得られた水抽出物からメタノール可溶物を除く工程、および
（ｉＶ）無機金属複合体を板状結晶として回収する工程、
からなるリモナイトから抗菌性無機複合体を分離する方法。
（１１）前記工程（ｉｉ）が、アセトン可溶物を除く工程、エーテル可溶物を除く工程、およびクロロホルム可溶物を除く工程のうちの少なくとも一つを含む、上記（１０）に記載の抗菌性無機金属複合体の分離方法。
（１２）前記工程（ｉｉ）が、アセトン可溶物を除く工程、エーテル可溶物を除く工程、及びクロロホルム可溶物を除く工程を含む、上記（１１）に記載の分離方法。
（１３）前記リモナイトが、リモナイトＬＭＢ３００である、上記（１０）〜（１２）のいずれか一つに記載の抗菌性無機金属複合体の分離方法。
（１４）上記（１）〜（５）のいずれか一つに記載の抗菌性無機金属複合体を含む抗菌剤。
(10) The following steps:
(I) suspending limonite in water and recovering the water extract;
(Ii) removing organic solvent solubles from the water extract;
(Iii) a step of removing methanol-soluble matter from the water extract obtained in the step (ii), and (iV) a step of recovering the inorganic metal composite as a plate-like crystal,
A method for separating an antibacterial inorganic complex from limonite comprising:
(11) The step (ii) includes at least one of a step of removing acetone-soluble matter, a step of removing ether-soluble matter, and a step of removing chloroform-soluble matter, according to (10) above. Method for separating antibacterial inorganic metal composite.
(12) The separation method according to (11), wherein the step (ii) includes a step of removing acetone-soluble matter, a step of removing ether-soluble matter, and a step of removing chloroform-soluble matter.
(13) The method for separating an antibacterial inorganic metal complex according to any one of (10) to (12), wherein the limonite is limonite LMB300.
(14) An antibacterial agent comprising the antibacterial inorganic metal composite according to any one of (1) to (5) above.
本発明により、新規な土壌由来の抗菌活性を有する無機金属複合体が提供された。そのような抗菌性無機金属複合体は、抗菌剤として有用である。 According to the present invention, a novel inorganic metal complex having antibacterial activity derived from soil was provided. Such an antibacterial inorganic metal composite is useful as an antibacterial agent.
以下、本発明を、例示的な実施態様を例として、本発明の実施において使用することができる好ましい方法および材料とともに説明する。なお、文中で特に断らない限り、本明細書で用いるすべての技術用語及び科学用語は、本発明が属する技術分野の当業者に一般に理解されるのと同じ意味をもつ。また、本明細書に記載されたものと同等または同様の任意の材料および方法は、本発明の実施において同様に使用することができる。 The invention will now be described by way of example embodiments, together with preferred methods and materials that can be used in the practice of the invention. Unless otherwise noted in the text, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition, any materials and methods equivalent or similar to those described herein can be used as well in the practice of the invention.
本発明の一つの態様は、原材料として、阿蘇・狩尾一帯に分布している「阿蘇の黄土」を採掘後、３年間熟成させた土壌であるリモナイト（株式会社日本リモナイト、ＬＭＢ３００）を用いて、抗菌性無機金属複合体を分離することである。
本発明の他の一つの態様は、リモナイトより分離された抗菌性無機金属複合体である。
本発明の別の一つの態様は、主要な金属としてＡｌ，Ｍｎ，Ｚｎ，Ｃａを含み、それらの金属を含む硫化塩とフッ化物の混合物であることを特徴とする抗菌性無機金属複合体である。
本発明の別の一つの態様は、下記式：
［Ａｌ，Ｍｎ，Ｃａ，ＳＯ4，Ｆ］・ｎＨ2Ｏ
で表される抗菌性無機金属複合体である。
One aspect of the present invention uses, as a raw material, limonite (Nihon Limonite Co., Ltd., LMB300), which is a soil that has been aged for 3 years after mining "Aso's loess" distributed in the Aso / Kao area. It is to separate the antibacterial inorganic metal complex.
Another embodiment of the present invention is an antibacterial inorganic metal composite separated from limonite.
Another aspect of the present invention is an antibacterial inorganic metal composite comprising Al, Mn, Zn, Ca as main metals and a mixture of sulfides and fluorides containing these metals. is there.
Another embodiment of the present invention is a compound of the following formula:
[Al, Mn, Ca, SO 4 , F] · nH 2 O
It is an antibacterial inorganic metal composite represented by.
土壌リモナイトからの抗菌性無機金属複合体の分離は、これに限定されないが、例えば、以下のようにして行うことができる。
土壌ＬＭＢ３００に水を加え、常温で数時間、例えば２時間震盪した後、水抽出液を回収し、次いで、濾過し、濃縮を行う。濃縮は、その後の操作が有機溶媒による抽出であるので、濃縮乾固を行うのが好ましい。有機溶媒の抽出は、これに限定されないが、例えば、アセトン抽出、エーテル抽出、クロロホルム抽出を、その順番にて行うことができる。本発明の抗菌性無機金属複合体は有機溶媒に溶解しないので、水抽出液を濃縮乾固した場合は、それぞれの有機溶媒による抽出工程において、不溶性部分（残渣）を回収し、次の有機溶媒による抽出を行い、常に残渣を回収することにより行うことができる。また、水抽出液を乾固させずに濃縮した場合は、それぞれの有機溶媒による抽出工程において、水溶性分画を回収し（つまり、有機溶媒分画を除去し）、次の有機溶媒による抽出を行い、常に水溶性分画を回収することにより行うことができる。その後、例えば、ゲルろ過クロマトグラフィー等を用いて不純物を除去することができる。次いで、不純物を除去した分画にメタノールを添加してメタノール可溶部分を除去して、本発明の抗菌性無機金属複合体を得ることができる。このようにして得られた複合体は、白色板状結晶であり、土壌リモナイトＬＭＢ３００の４００ｇ中に１ｇ以上含まれている。
The separation of the antibacterial inorganic metal complex from the soil limonite is not limited to this, but can be performed, for example, as follows.
After adding water to the soil LMB300 and shaking at room temperature for several hours, for example, 2 hours, the water extract is recovered, then filtered and concentrated. Concentration is preferably carried out by concentration and drying since the subsequent operation is extraction with an organic solvent. Although extraction of an organic solvent is not limited to this, For example, acetone extraction, ether extraction, and chloroform extraction can be performed in the order. Since the antibacterial inorganic metal complex of the present invention is not dissolved in an organic solvent, when the water extract is concentrated to dryness, insoluble portions (residues) are recovered in the extraction step with each organic solvent, and the following organic solvent The extraction can be performed by always collecting the residue. In addition, when the water extract is concentrated without drying, the water-soluble fraction is recovered (that is, the organic solvent fraction is removed) in the extraction step with each organic solvent and extracted with the next organic solvent. And always collecting the water-soluble fraction. Thereafter, for example, impurities can be removed using gel filtration chromatography or the like. Subsequently, methanol is added to the fraction from which impurities have been removed to remove the methanol-soluble portion, whereby the antibacterial inorganic metal composite of the present invention can be obtained. The complex thus obtained is a white plate crystal and is contained in 1 g or more in 400 g of soil limonite LMB300.
本発明の抗菌性無機金属複合体（以下、単に「本発明の複合体」という場合がある）は、主要な金属としてＡｌ，Ｍｎ，Ｚｎ，Ｃａを含み、それらの金属を含む硫化塩とフッ化物の混合物であることを特徴としている。
本発明の複合体は、それに含まれる金属の含有量は、Ａｌ＞Ｍｎ＞Ｚｎ，Ｃａであることを特徴とする。
本発明の複合体はまた、さらに他の金属を含んでもよい。これに限定されないが、例えば、Ｎａをあげあることができ、係る金属は、Ｚｎ，Ｃａより少ない量で含まれる。
The antibacterial inorganic metal composite of the present invention (hereinafter sometimes simply referred to as “composite of the present invention”) contains Al, Mn, Zn, Ca as main metals, and sulfides and fluorides containing these metals. It is characterized by being a mixture of compounds.
The composite of the present invention is characterized in that the content of the metal contained therein is Al>Mn> Zn, Ca.
The composite of the present invention may further contain other metals. Although it is not limited to this, for example, Na can be mentioned and such a metal is contained in a smaller amount than Zn and Ca.
本発明の複合体は、水溶性で、有機溶媒に不溶性であることを特徴とする。
本発明の複合体は、グラム陽性菌（例えば、B.subtilis）、グラム陰性菌（例えば、Ps.aeruginosa）、および真菌（例えば、Candida albicans）のいずれにも強い抗菌作用をもち、それは明礬の抗菌性よりも強い、という特徴をもつ。
本発明の複合体は、グラム陰性菌、グラム陽性菌、真菌のいずれの菌に対しても、ＭＩＣ値で、２ｍｇ／ｍｌ以下、好ましくは１ｍｇ／ｍｌ以下、例えば、０．５〜１ｍｇ／ｍｌで抗菌、抗真菌活性を示す。
本発明の複合体は、以下の式：
［Ａｌ，Ｍｎ，Ｃａ，ＳＯ4，Ｆ］・ｎＨ2Ｏ
（ｎは正数である）であらわされる、硫化塩およびフッ化物を含む複塩である。
The complex of the present invention is water-soluble and insoluble in an organic solvent.
The complex of the present invention has a strong antibacterial action against any of Gram-positive bacteria (eg, B. subtilis), Gram-negative bacteria (eg, Ps. Aeruginosa), and fungi (eg, Candida albicans). It is characterized by stronger than antibacterial properties.
The complex of the present invention has a MIC value of 2 mg / ml or less, preferably 1 mg / ml or less, for example 0.5 to 1 mg / ml, against any of Gram-negative bacteria, Gram-positive bacteria and fungi. With antibacterial and antifungal activity.
The complex of the present invention has the following formula:
[Al, Mn, Ca, SO 4 , F] · nH 2 O
(N is a positive number) and is a double salt containing sulfide and fluoride.
以下、本発明を、以下に記載の実施例をもとに説明するが、本発明は以下の実施例に限定されるものではない。 EXAMPLES Hereinafter, although this invention is demonstrated based on the Example described below, this invention is not limited to a following example.
（１）材料および方法
阿蘇・狩尾一帯に分布している「阿蘇の黄土」を採掘後、３年間熟成させた土壌リモナイトＬＭＢ３００（株式会社日本リモナイト、ＬＭＢ３００）を用いた。
（２）抗菌活性の測定
抗菌活性の測定は、ペーパーディスク法および希釈法により行った。
培地はブイヨン寒天培地またはグルコースブイヨン寒天培地を用いた。
ペーパーディスク法では、試験菌として、Bacillus subtilisを用いた。
希釈法では、寒天希釈法を用い、試験菌としては、Bacilus subtillis, Staphylococcus aureus, Micrococcus luteus, Escherichia coli, Proteus vulgaris, Psudomonas aeruginosa, Serratia marcescence, Candida albicans,および Aspergillus nigerを用いた。
(1) Materials and Methods Soil limonite LMB300 (Nihon Limonite Co., Ltd., LMB300) aged for 3 years after mining “Aso loess” distributed in the Aso / Kario area was used.
(2) Measurement of antibacterial activity The antibacterial activity was measured by a paper disk method and a dilution method.
As the medium, a bouillon agar medium or a glucose bouillon agar medium was used.
In the paper disk method, Bacillus subtilis was used as a test bacterium.
In the dilution method, an agar dilution method was used, and Bacilus subtillis, Staphylococcus aureus, Micrococcus luteus, Escherichia coli, Proteus vulgaris, Psudomonas aeruginosa, Serratia marcescence, Candida albicans, and Aspergillus niger were used as test bacteria.
（実施例１）土壌リモナイト中の抗菌成分の確認
本発明者らは先ず始めに、土壌リモナイトＬＭＢ３００中に抗菌成分が含まれるかどうかを確認するため、水、メタノール、熱水（９０℃）、もしくは熱メタノール（６２℃）により抽出操作を行い、抽出液の抗菌活性をディスク法にて検定した。その結果、すべての抽出条件で抽出物に抗菌力が認められ、ＬＭＢ３００に抗菌成分が含まれることが確認できた。
そこで、以下のようにして、土壌リモナイトＬＭＢ３００から抗菌性成分を分離した。
(Example 1) Confirmation of antibacterial component in soil limonite First, in order to confirm whether the antibacterial component is contained in the soil limonite LMB300, the present inventors, water, methanol, hot water (90 ° C), Alternatively, extraction was performed with hot methanol (62 ° C.), and the antibacterial activity of the extract was assayed by the disk method. As a result, antibacterial activity was recognized in the extract under all extraction conditions, and it was confirmed that the antibacterial component was contained in LMB300.
Therefore, antibacterial components were separated from the soil limonite LMB300 as follows.
（実施例２）抗菌性成分の分離
土壌リモナイトＬＭＢ３００の５０ｇに水２００ｍＬを加え、常温で２時間震盪し、濾過、濃縮、乾固した後、適量のアセトンを加えその残渣を得た。その残渣に適量のエーテルを加えその残渣を得た。さらにその残渣に適量のクロロホルムを加え残渣を得た。その残渣に水１０ｍＬを加え水溶性成分を含む水分画を得た。得られた水分画をＳｅｐｈａｄｅｘ Ｇ−５０ カラムクロマトグラフィー（ＧＥヘルスケア社）に供した。各フラクションの抗菌活性を測定した。抗菌活性は、ペーパーディスク法により確認した。抗菌活性を有するフラクションは９〜１１フラクションであった。この抗菌活性を有するフラクションを活性炭クロマトグラフィー（和光純薬株式会社）に供し脱色操作を行った。その後さらにＳｅｐｈａｄｅｘ Ｇ−１０ カラムクロマトグラフィー（ＧＥヘルスケア社）に供し抗菌活性を示す７〜１１フラクションを分離した。このフラクションを濃縮、乾固し、適量のメタノールを加えてメタノール可溶部分を除いて白色板状結晶を得た。この白色板状結晶の最終産物をＬＭＢ−Ｗと称す。ＬＭＢ−Ｗは４００ｇの土壌ＬＭＢ３００の中に１ｇ以上含まれていた。これらの工程の概略を図１に示す。
(Example 2) Separation of antibacterial component After adding 200 mL of water to 50 g of soil limonite LMB300, shaking at room temperature for 2 hours, filtering, concentrating and drying, an appropriate amount of acetone was added to obtain a residue. An appropriate amount of ether was added to the residue to obtain the residue. Further, an appropriate amount of chloroform was added to the residue to obtain a residue. 10 mL of water was added to the residue to obtain a water fraction containing a water-soluble component. The obtained water fraction was subjected to Sephadex G-50 column chromatography (GE Healthcare). The antibacterial activity of each fraction was measured. Antibacterial activity was confirmed by the paper disk method. Fractions having antibacterial activity were 9 to 11 fractions. The fraction having antibacterial activity was subjected to activated carbon chromatography (Wako Pure Chemical Industries, Ltd.) for decolorization operation. Thereafter, it was further subjected to Sephadex G-10 column chromatography (GE Healthcare) to separate 7 to 11 fractions exhibiting antibacterial activity. This fraction was concentrated and dried, and an appropriate amount of methanol was added to remove the methanol-soluble portion to obtain white plate crystals. The final product of this white plate crystal is referred to as LMB-W. LMB-W was contained in 1 g or more in 400 g of soil LMB300. An outline of these steps is shown in FIG.
（実施例３）ＬＭＢ−Ｗの抗菌、抗真菌活性
実施例２で得られたＬＭＢ−Ｗの抗菌、抗真菌活性を、寒天希釈法により測定した。その結果を表１に示す。ＬＭＢ−Ｗはグラム陽性菌、グラム陰性菌、および真菌に対して強い抗菌力を示した。
(Example 3) Antibacterial and antifungal activity of LMB-W The antibacterial and antifungal activity of LMB-W obtained in Example 2 was measured by an agar dilution method. The results are shown in Table 1. LMB-W showed strong antibacterial activity against gram positive bacteria, gram negative bacteria and fungi.
（実施例４）他の抗菌性金属複合体との比較
希釈法を用いて、ＬＭＢ−Ｗの抗菌・抗真菌活性を、ミョウバン類およびヒューマス・カルミーから抽出した抗菌性成分（ＫＨ−Ｃ1）と比較した。結果を表２に示す。ＬＭＢ−Ｗの抗菌・抗真菌活性はミョウバン類と比較すると、グラム陽性菌（B.subtilis）、グラム陰性菌（Ps.aeruginosa）、真菌（Candida albicans）それぞれに対して約５倍の活性を示した。ＬＭＢ−ＷをＫＨ−Ｃ1と比較すると、グラム陽性菌に対して約５倍、グラム陰性菌に対して約２倍、および真菌に対しては同等の抗菌・抗真菌活性を示した。
(Example 4) An antibacterial component (KH-C 1 ) obtained by extracting the antibacterial / antifungal activity of LMB-W from alums and human carme using a comparative dilution method with other antibacterial metal complexes Compared with. The results are shown in Table 2. The antibacterial and antifungal activities of LMB-W are about 5 times higher than that of alums against Gram-positive bacteria (B. subtilis), Gram-negative bacteria (Ps. Aeruginosa) and fungi (Candida albicans). It was. When the LMB-W compared to KH-C 1, about 5-fold against gram-positive bacteria, twice against gram-negative bacteria, and against fungi showed comparable antibacterial and antifungal activity.
（実施例５）ＬＭＢ−Ｗの化学組成分析
ＬＭＢ−Ｗの化学組成を、ＩＣＰ−ＭＳ（ＥＬＥＭＥＮＴ、サーモフィッシャーサイエンティフィック社）、ＩＣＰ発光原子吸光分析（Ｈｉ−ＣＡＰ６３００、サイエンティフィック社）、および陰イオンカラムクロマトグラフィー（ウォーターズ４３１、カラムＩＣ−ＰＡＣＡ、ウォーターズ社）を用いて行った。検討結果を以下の表３に示す。このように、ＬＭＢ−Ｗは、元素としてＣとＮを含まないことが確認できた。また陰イオンカラムクロマトグラフィーより、ＳＯ4 2-およびＦ-が存在することが確認できた。その結果、ＬＭＢ−Ｗは、明礬類やＫＨ−Ｃ1とは異なる化学組成を示しており、マンガンとフッ素を含むことを特徴とする無機金属の複合体であることが判った。
(Example 5) Chemical composition analysis of LMB-W The chemical composition of LMB-W was measured using ICP-MS (ELEMENT, Thermo Fisher Scientific), ICP emission atomic absorption spectrometry (Hi-CAP 6300, Scientific), And anion column chromatography (Waters 431, Column IC-PACA, Waters). The examination results are shown in Table 3 below. Thus, it was confirmed that LMB-W does not contain C and N as elements. In addition, it was confirmed from the anion column chromatography that SO 4 2− and F − were present. As a result, LMB-W shows a different chemical composition than the alum compound and KH-C 1, was found to be a composite of inorganic metal, characterized in that it contains manganese and fluorine.
（実施例６）ＬＭＢ−Ｗの化学組成分析
さらに、実施例２により分離・精製したＬＭＢ−Ｗを結晶化し、蛍光Ｘ線分析（ＳＩＩ製ＳＥＡ２２２０Ａ）を行った。結果を以下の表４に示す。これにより、ＬＭＢ−Ｗを構成する主要な金属は、Ａｌ，Ｍｎ，Ｚｎ，およびＣａであることが判った。
(Example 6) Chemical composition analysis of LMB-W Further, LMB-W separated and purified in Example 2 was crystallized and subjected to X-ray fluorescence analysis (SEA2220A manufactured by SII). The results are shown in Table 4 below. Thereby, it turned out that the main metals which comprise LMB-W are Al, Mn, Zn, and Ca.
上記の詳細な記載は、本発明の目的及び対象を単に説明するものであり、添付の特許請求の範囲を限定するものではない。添付の特許請求の範囲から離れることなしに、記載された実施態様に対しての、種々の変更及び置換は、本明細書に記載された教示より当業者にとって明らかである。 The above detailed description is merely illustrative of the objects and objects of the invention and is not intended to limit the scope of the appended claims. Various changes and substitutions to the described embodiments will be apparent to those skilled in the art from the teachings described herein without departing from the scope of the appended claims.
本発明により提供された、新規な土壌由来の抗菌活性を有する無機金属複合体は、グラム陽性菌、グラム陰性菌および真菌に対して優れた抗菌活性を有し、抗菌剤として有用である。 The novel inorganic metal complex having antibacterial activity derived from soil provided by the present invention has excellent antibacterial activity against gram-positive bacteria, gram-negative bacteria and fungi, and is useful as an antibacterial agent.
Claims (14)
（ａ）主要な金属として、Ａｌ，Ｍｎ，Ｚｎ，およびＣａを含む、
（ｂ）前記金属の含有量は、Ａｌ＞Ｍｎ＞Ｚｎ，Ｃａである。および
（ｃ）グラム陽性菌、グラム陰性菌および真菌に対して抗菌活性をもつ、
の特徴を有する抗菌性無機金属複合体。 following:
(A) including Al, Mn, Zn, and Ca as main metals,
(B) The metal content is Al>Mn> Zn, Ca. And (c) has antibacterial activity against gram positive bacteria, gram negative bacteria and fungi,
An antibacterial inorganic metal composite having the following characteristics:
［Ａｌ，Ｍｎ，Ｃａ，ＳＯ４，Ｆ］・ｎＨ２Ｏ
であらわされる、硫化塩およびフッ化物を含む複塩である、請求項１に記載の抗菌性無機金属複合体。 formula:
[Al, Mn, Ca, SO 4 , F] · nH 2 O
The antibacterial inorganic metal composite according to claim 1, which is a double salt containing a sulfide and a fluoride.
（ｉ）リモナイトを水に懸濁し、水抽出物を回収する工程、
（ｉｉ）前記水抽出物から、有機溶媒可溶物を除く工程、
（ｉｉｉ）上記工程（ｉｉ）により得られた水抽出物からメタノール可溶物を除く工程、
により抽出される抗菌性無機金属複合体。 The following steps from limonite:
(I) suspending limonite in water and recovering the water extract;
(Ii) removing organic solvent solubles from the water extract;
(Iii) a step of removing methanol-soluble matter from the water extract obtained by the step (ii),
Antibacterial inorganic metal composites extracted by
（ｉ）リモナイトを水に懸濁し、水抽出物を回収する工程、
（ｉｉ）前記水抽出物から、有機溶媒可溶物を除く工程、
（ｉｉｉ）上記工程（ｉｉ）により得られた水抽出物からメタノール可溶物を除く工程、および
（ｉＶ）無機金属複合体を板状結晶として回収する工程、
からなるリモナイトから抗菌性無機複合体を分離する方法。 The following steps,
(I) suspending limonite in water and recovering the water extract;
(Ii) removing organic solvent solubles from the water extract;
(Iii) a step of removing methanol-soluble matter from the water extract obtained in the step (ii), and (iV) a step of recovering the inorganic metal composite as a plate-like crystal,
A method for separating an antibacterial inorganic complex from limonite comprising:
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