New hydroxamate metabolite, MBJ-0003, from Micromonospora sp. 29867
New hydroxamate metabolite, MBJ-0003, from Micromonospora sp. 29867
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来自小单孢菌属的新异羟肟酸代谢物 MBJ-0003。
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
K. Shin‐ya
中科院分区：
文献类型：
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作者：
Teppei Kawahara;Masashi Itoh;M. Izumikawa;I. Kozone;Noriaki Sakata;T. Tsuchida;K. Shin‐ya
在筛选微生物新生物活性物质的过程中，我们使用先进的化合物鉴定系统“MBJ的特殊选择，1,2”，选择了50多个菌株培养样品，从而成功地从Nectria sp. f261113和Apiognomonia sp. f240231,1中分别发现了新的细胞毒性细菌衍生物MBJ-0009， MBJ-0010和MBJ-0011， MBJ-0012和MBJ-0013，以及新的毛球蛋白衍生物MBJ-0038。进一步筛选从小单孢菌sp. 29867培养物中分离出新的羟酸盐代谢物MBJ-0039和MBJ-0040（1，图1a）。本文介绍了菌株1的发酵、分离、结构鉴定和生物活性。小单孢子虫sp. 29867从日本静冈县骏河湾采集的贝类中分离得到。菌株在250ml的Erlenmeyer烧瓶中培养，每个烧瓶含有25ml的培养基，培养基由2%马铃薯淀粉（Tobu Tokachi Nosan Kako农业合作协会，日本北海道）、2%葡萄糖（Junsei Chemical，日本东京）、2%大豆粉（Honen SoyPro, J-Oil Mills，日本东京）、0.5%酵母提取物粉（Oriental yeast，日本东京）、0.25% NaCl （Junsei Chemical）、0.32% CaCO3 （Wako Pure Chemical Industries，日本大阪）、0.0005% CuSO4 5H2O (Wako Pure Chemical Industries), 0.0005% ZnSO4.7H2O （Wako Pure Chemical Industries）和0.0005% MnCl2 4H2O （Junsei Chemical）。将烧瓶置于28℃的旋转摇床（220转/分）上，孵育3天。然后将等份（0.5ml）的肉汤转移到500ml的Erlenmeyer烧瓶中，其中含有50ml相同的培养基，并在28℃的旋转摇床（220转/分）上培养4天。用等体积的正丁醇提取整个培养液（2 l）。将正丁醇层真空蒸发至干燥，残渣依次悬浮于水中（350ml），然后用乙酸乙酯（350ml 3）洗涤，再用正丁醇（300ml 2）萃取。正丁醇层蒸发后，提取液（2.9 g）采用正相中压液相色谱法（Purif-Pack SI-30，尺寸：60 (29 g), Shoko Scientific, Yokohama，日本），用正己烷-乙酸乙酯梯度体系（0 - 25%乙酸乙酯，12 min， 25%保持3min，流速：20ml min - 1），然后用CHCl3-MeOH溶剂体系（0、2、5、10、20、30和100% MeOH各100ml）逐步开发。采用UPLC-DAD-ELS-MS系统对各组分进行监测。100% MeOH洗脱部分（608mg）采用反相中压液相色谱法（purf - pak ODS-30，尺寸：60 (30 g), Shoko Scientific），用H2O-MeOH-CHCl3体系的显像溶剂（60、70、80和100%水溶液MeOH和CHCl3/MeOH1 / 4 1:1各100ml）进行色谱。CHCl3/MeOH洗脱液（214mg）在CAPCELL PAK C18 MGII色谱柱（5.0mm, 20 i.d. 150mm; Shiseido, Tokyo, Japan）上采用制备高效液相色谱法（HPLC）， 55%水溶液中含有0.1%甲酸（流速：10ml min-1），得到半纯化1 （7.9mg，保留时间：9.0min）。最终纯化采用制备高效液相色谱法，采用XSelect CSH C18色谱柱（20 id . 150mm; Waters, Milford， MA， USA）， 40%水溶液中含有0.1%甲酸（流速：10ml min-1），得到1 （1.7mg，保留时间：16.3min）。1是一种非旋光性化合物；一种无色无定形固体，具有紫外端和红外（ATR） nmax 3300和1650 cm 1（羟基和羰基）。采用负模式高分辨率电喷雾电离质谱法（m/z 727.5353 [m - h] -，计算C37H71N6O8 727.5333）建立分子式为C37H72N6O8。质子和碳之间的直接连通性是由异核单量子相干谱建立的；表1列出了1的13C和1H核磁共振光谱数据。通过分析DQF-COSY和恒时异核多键相关（CT-HMBC）4数据进行1的结构确定，如下所述。DQF-COSY和CT-HMBC光谱显示有7个片段（A到G），如下图（图1b）。脂肪族之间的1H自旋偶联揭示了尸胺片段（片段C）
In the course of our screening program for new bioactive substances from microorganisms using the advanced compound-identification system designated as ‘MBJ’s special selection,1,2 with which we have selected over 50 strain culture samples and consequently have succeeded in discovering new cytotoxic eremophilane derivatives MBJ-0009, MBJ-0010 and MBJ-0011, MBJ-0012 and MBJ-0013 from Nectria sp. f261113 and Apiognomonia sp. f24023,1 respectively, and new chaetoglobosin derivatives MBJ-0038, MBJ-0039 and MBJ-0040 from Chaetomium sp. f24230.2 Further screening resulted in the isolation of a new hydroxamate metabolite, designated as MBJ-0003 (1, Figure 1a), from the culture of Micromonospora sp. 29867. In this paper, we described the fermentation, isolation, structure elucidation and brief biological activities of 1. Micromonospora sp. 29867 was isolated from a shellfish collected in Suruga Bay, Shizuoka Prefecture, Japan. The strain was cultivated in 250ml Erlenmeyer flasks, each containing 25ml of a medium consisting of 2% potato starch (Tobu Tokachi Nosan Kako Agricultural Cooperative Association, Hokkaido, Japan), 2% glucose (Junsei Chemical, Tokyo, Japan), 2% soy bean powder (Honen SoyPro, J-Oil Mills, Tokyo, Japan), 0.5% yeast extract powder (Oriental Yeast, Tokyo, Japan), 0.25% NaCl (Junsei Chemical), 0.32% CaCO3 (Wako Pure Chemical Industries, Osaka, Japan), 0.0005% CuSO4 5H2O (Wako Pure Chemical Industries), 0.0005% ZnSO4.7H2O (Wako Pure Chemical Industries) and 0.0005% MnCl2 4H2O (Junsei Chemical). The flasks were placed on a rotary shaker (220 r.p.m.) at 28 1C and incubated for 3 days. Aliquots (0.5ml) of the broth were then transferred to 500ml Erlenmeyer flasks containing 50ml of the same medium and were cultured on a rotary shaker (220 r.p.m.) at 28 1C for 4 days. The whole culture broth (2 l) was extracted with an equal volume of n-BuOH. The n-BuOH layer was evaporated in vacuo to dryness and the residue was successively suspended in water (350ml), then washed with EtOAc (350ml 3), followed by extraction with n-BuOH (300ml 2). The n-BuOH layer was evaporated and the extract (2.9 g) was subjected to normal-phase medium-pressure liquid chromatography (Purif-Pack SI-30, size: 60 (29 g), Shoko Scientific, Yokohama, Japan) and developed with a gradient system of n-hexane– EtOAc (0–25% EtOAc over 12 min and was kept at 25% for 3min, flow rate: 20ml min–1), followed by the stepwise solvent system of CHCl3–MeOH (100ml each of 0, 2, 5, 10, 20, 30 and 100% MeOH). The fractions were monitored by using an UPLC-DAD-ELS-MS system. The 100% MeOH-eluted fraction (608mg) was then chromatographed by reversed-phase medium-pressure liquid chromatography (Purif-Pak ODS-30, size: 60 (30 g), Shoko Scientific) with the developing solvent of the H2O–MeOH–CHCl3 system (100ml each of 60, 70, 80 and 100% aqueous MeOH and CHCl3/MeOH1⁄4 1:1). The CHCl3/MeOH eluate (214mg) was subjected to preparative HPLC on a CAPCELL PAK C18 MGII column (5.0mm, 20 i.d. 150mm; Shiseido, Tokyo, Japan) with 55% aqueous MeCN containing 0.1% formic acid (flow rate: 10ml min–1) to give semi-purified 1 (7.9mg; retention time: 9.0min). Final purification was carried out by preparative HPLC on an XSelect CSH C18 column (20 i.d. 150mm; Waters, Milford, MA, USA) with 40% aqueous MeCN containing 0.1% formic acid (flow rate: 10ml min–1) to afford 1 (1.7mg; retention time: 16.3min). 1 was found to be an optically inactive compound; a colorless amorphous solid with UV end and IR (ATR) nmax 3300 and 1650 cm 1 (hydroxy and carbonyl). The molecular formula was established as C37H72N6O8 by negative mode high-resolution electrospray ionization mass spectrometry (m/z 727.5353 [M–H]–, calcd for C37H71N6O8 727.5333). The direct connectivity between protons and carbons was established by a heteronuclear single-quantum coherence spectrum; the tabulated 13C and 1H NMR spectroscopic data for 1 are listed in Table 1. The structure determination of 1 was carried out through analysis of DQF-COSY and constant-time heteronuclear multiple bond correlation (CT-HMBC)4 data as described below. DQF-COSY and CT-HMBC spectra revealed the presence of seven fragments (A to G) as follows (Figure 1b). A cadaverine moiety (fragment C) was revealed by the 1H spin couplings between aliphatic
影响因子：
16.6
作者：
Yang, Yu-Liang;Xu, Yuquan;Kersten, Roland D.;Liu, Wei-Ting;Meehan, Michael J.;Moore, Bradley S.;Bandeira, Nuno;Dorrestein, Pieter C.
通讯作者：
Dorrestein, Pieter C.