王鳴慧, 李曉明 李春順 王斌貴

(1. 中國科學(xué)院 海洋研究所 實(shí)驗(yàn)海洋生物學(xué)重點(diǎn)實(shí)驗(yàn)室, 山東 青島 266071; 2. 中國科學(xué)院大學(xué), 北京 100049)

海洋沉積物來源真菌嗜松青霉SD-272代謝產(chǎn)物研究

王鳴慧1,2, 李曉明1, 李春順1, 王斌貴1

(1. 中國科學(xué)院 海洋研究所 實(shí)驗(yàn)海洋生物學(xué)重點(diǎn)實(shí)驗(yàn)室, 山東 青島 266071; 2. 中國科學(xué)院大學(xué), 北京 100049)

對來源于珠江口沉積物的一株海洋真菌嗜松青霉(Penicillium pinophilum)SD-272的次生代謝產(chǎn)物, 進(jìn)行了化學(xué)成分分離及其生物活性研究。采用常規(guī)硅膠柱層析、凝膠Sephadex LH-20柱層析、制備薄層層析等分離手段, 通過紫外、核磁共振技術(shù)、質(zhì)譜技術(shù)等現(xiàn)代波譜學(xué)技術(shù), 從其發(fā)酵液提取物中共分離并鑒定了 13個(gè)化合物, 分別為: 4′-demethylvermistatin (1), vermistatin (2), penisimplicissin(3), deoxyfunicone (4), 5, 6-epoxy-3-deoxyfunicone (5), 5′-methoxy-6-methyl-biphenyl-3, 4, 3′-triol (6), altenusin (7), 1-deoxyrubralactone(8), kojic acid (9), 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (10), dankasterone (11), 4-hydroxy-2-methoxyacetanilide (12), N-(2-hydroxypropanoyl)-2-aminobenzoic acid amide (13)。這些化合物均為首次從嗜松青霉中分離得到, 其中化合物11顯示較好的鹵蟲致死活性, LD50值為39.2 μmol/L。

沉積物; 嗜松青霉(Penicillium pinophilum); 次生代謝產(chǎn)物; 鹵蟲致死活性

海洋沉積物是在漫長的地質(zhì)年代里, 由陸地徑流、大氣輸入及人類活動帶入海洋并沉降在海底的物質(zhì)總稱。海洋沉積物既不同于淡水沉積物和土壤等陸地環(huán)境, 又與海水環(huán)境相對獨(dú)立, 長期處于高鹽、低溫和高壓等特殊生態(tài)環(huán)境, 成為地球上最復(fù)雜的微生物棲息地[1-2]。海洋沉積物來源真菌作為海洋微生物重要組成部分, 蘊(yùn)藏著許多具有潛在應(yīng)用前景的活性物質(zhì), 是開發(fā)新抗菌、抗腫瘤藥物等新藥的重要資源[3]。近年來, 科學(xué)研究已在海洋沉積物來源真菌次生代謝產(chǎn)物中發(fā)現(xiàn)了大量結(jié)構(gòu)新穎、活性多樣的化合物[4-7]。在本課題組對海底沉積物來源真菌次生代謝產(chǎn)物研究的過程中[8-9], 我們對一株珠江口沉積物來源的真菌嗜松青霉(Penicillium pinophilum)進(jìn)行了規(guī)模發(fā)酵, 從其發(fā)酵提取物中分離鑒定了 13個(gè)化合物(圖 1), 這些化合物均為首次從嗜松青霉中分離得到, 其中化合物11顯示較好的鹵蟲致死活性, LD50值為39.2 mol/L。

1 材料與方法

1.1 儀器與試劑

BrukerAvance 500 MHz核磁共振儀; Dionex分析型高效液相色譜儀; Lobar LiChroprer RP-18硅膠(40～63 μm, Merck); 薄層色譜硅膠GF254和柱色譜硅膠(200～300目)為青島海洋化工分廠產(chǎn)品; 顯色劑為茴香醛硫酸溶液和碘; 所有有機(jī)溶劑均為重蒸工業(yè)級溶劑。

1.2 菌株發(fā)酵

(1) 菌株: 菌株 SD-272是分離自珠江口沉積物中的真菌。

(2) 菌株發(fā)酵: 菌種以瓊脂-麥芽膏培養(yǎng)基 4℃保存。發(fā)酵培養(yǎng)基成分為: 蔗糖2%、蛋白胨0.5%、酵母膏0.3%、味精1%、甘露醇2%、土豆汁20%, 培養(yǎng)基pH為6.5。盛有發(fā)酵培養(yǎng)基的1 L三角瓶(60瓶)在 116℃下高壓滅菌 20 min, 待三角瓶冷卻后接種,常溫靜置培養(yǎng)35 d。

1.3 提取分離

菌株規(guī)模發(fā)酵結(jié)束后, 發(fā)酵液采用乙酸乙酯萃取, 減壓蒸干得到發(fā)酵液提取物; 菌絲體經(jīng)細(xì)胞破碎儀破碎后, 采用80%丙酮水溶液超聲萃取3次, 減壓濃縮去除丙酮后的剩余水相再用乙酸乙酯萃取 3次, 減壓蒸干有機(jī)相得到菌絲體粗提物, 經(jīng)HPLC和 TLC檢測發(fā)現(xiàn)與發(fā)酵液提取物基本一致, 合并后得到總粗提物50.2 g。

將上述粗提物進(jìn)行硅膠VLC柱層析, 根據(jù)極性從小到大(石油醚/乙酸乙酯到氯仿/甲醇)進(jìn)行梯度洗脫, 經(jīng) TLC和 HPLC檢測, 合并得到 9個(gè)組分(Fr.1–9)。其中, Fr.2經(jīng)正相硅膠柱層析、凝膠Sephadex LH-20(甲醇)柱層析和制備薄層層析分離得到化合物1(6.3 mg)、2(25.1 mg)、3(5.8 mg)、4(10.1 mg)和化合物 5(6.7 mg); Fr.3經(jīng)反相硅膠柱層析、凝膠Sephadex LH-20(甲醇)柱層析和制備薄層層析分離得到化合物6(9.4 mg)、7(14.8 mg)、8(5.2 mg)和化合物11(8.1 mg); Fr.4經(jīng)正相硅膠柱層析、凝膠Sephadex LH-20(丙酮)柱層析、制備薄層層析和制備高效液相分離得到化合物9(6.3 mg)、10(3.1 mg)、12(6.3 mg)和化合物13(5.5 mg)。

圖1 化合物1–13的結(jié)構(gòu)Fig.1 Structures of compounds 1–13

1.4 鹵蟲致死活性篩選

(1) 材料: 鹵蟲卵取自中國科學(xué)院煙臺海岸帶研究所。

(2) 鹵蟲致死活性篩選[10]: 陽性對照藥為秋水仙堿, 陰性對照為DMSO, 空白對照為潔凈海水。取適量蟲卵放入潔凈海水中, 在室溫下通氣培養(yǎng)48 min后, 吸取195 μL鹵蟲生長液依次加入96孔板中, 再依次加入5 μL濃度為4 mg/mL的樣品溶液,使樣品終濃度達(dá)到0.1 mg/mL, 對每個(gè)樣品分別做3組平行實(shí)驗(yàn)。24 h后在解剖鏡下記錄每孔鹵蟲死亡數(shù)目和鹵蟲總數(shù), 計(jì)算致死率。

致死率 = 鹵蟲死亡數(shù)目/鹵蟲總數(shù) × 100%

2 化合物結(jié)構(gòu)鑒定

化合物1: 白色無定形粉末,1H-NMR (DMSO-d6)δH: 6.22 (1H, s, H-3), 8.14 (1H, s, H-6), 6.26 (1H, dd,J= 15.8, 1.6 Hz, H-7), 6.63 (1H, m, H-8), 1.88 (3H, dd,J= 6.9, 1.5 Hz, H-9), 6.22 (1H, s, H-10), 6.74 (1H, d,J= 1.6 Hz, H-3'), 6.73 (1H, d,J= 1.6 Hz, H-5'), 3.73 (3H, s, H-8');13C NMR (DMSO-d6)δC: 161.7 (C-2, C), 112.3 (C-3, CH), 176.3 (C-4, C), 122.4 (C-5, C), 155.5 (C-6, CH), 122.9 (C-7, CH), 135.8 (C-8, CH), 18.1 (C-9, CH3), 74.2 (C-10, CH), 124.7 (C-1', C), 128.6 (C-2', C), 101.8 (C-3', CH), 161.2 (C-4', C), 105.0 (C-5', CH), 154.7 (C-6', C), 169.8 (C-7', C), 55.7 (C-8', CH3)。其波譜數(shù)據(jù)與4'-demethylvermistatin的文獻(xiàn)報(bào)道[11]一致, 該化合物為首次從P.pinophilum中分離得到。當(dāng)樣品濃度為0.1 mg/mL時(shí), 對鹵蟲有一定的致死活性, 致死率為33%。

化合物2: 白色無定形粉末,1H-NMR (CDCl3)δH: 6.15 (1H, s, H-3), 7.42 (1H, s, H-6), 6.05 (1H, d,J= 15.0 Hz, H-7), 6.60 (1H, m, H-8), 1.91 (3H, d,J= 6.8 Hz, H-9), 6.44 (1H, s, H-10), 6.97 (1H, s, H-3'), 6.67 (1H, s, H-5'), 3.78 (3H, s, H-8'), 3.86 (3H, s, H-9');13C NMR (CDCl3)δC: 163.1 (C-2, C), 112.8 (C-3, CH), 177.2 (C-4, C), 123.4 (C-5, C), 153.8 (C-6, CH), 123.1 (C-7, CH), 135.9 (C-8, CH), 18.5 (C-9, CH3), 73.5 (C-10, CH), 127.7 (C-1', C), 129.3 (C-2', C), 99.0 (C-3', CH), 162.1 (C-4', C), 105.1 (C-5', CH), 154.9 (C-6', C), 169.9 (C-7', C), 55.7 (C-8', CH3), 55.9 (C-9', CH3)。其波譜數(shù)據(jù)與 vermistatin的文獻(xiàn)報(bào)道[12]一致, 該化合物為首次從P.pinophilum中分離得到。當(dāng)樣品濃度為0.1 mg/mL時(shí), 對鹵蟲有一定的致死活性, 致死率為41%。

化合物3: 白色無定形粉末,1H-NMR (DMSO-d6)δH: 6.17 (1H, s, H-3), 8.22 (1H, s, H-6), 1.75 (3H, s, H-7), 6.27 (1H, s, H-8), 6.92 (1H, d,J= 1.7 Hz, H-3'), 6.87 (1H, d,J= 1.7 Hz, H-5'), 3.86 (3H, s, H-8'), 3.76 (3H, s, H-9');13C NMR (DMSO-d6)δC: 166.5 (C-2, C), 114.4 (C-3, CH), 176.0 (C-4, C), 121.9 (C-5, C), 156.2 (C-6, CH), 18.9 (C-7, CH3), 74.3 (C-8, CH), 127.5 (C-1', C), 128.8 (C-2', C), 99.0 (C-3', CH), 162.3 (C-4', C), 104.9 (C-5', CH), 154.7 (C-6', C), 169.6 (C-7', C), 55.9 (C-8', CH3), 56.0 (C-9', CH3)。其波譜數(shù)據(jù)與penisimplicissin的文獻(xiàn)報(bào)道[12]一致, 該化合物為首次從P.pinophilum中分離得到。當(dāng)樣品濃度為0.1 mg/mL時(shí), 對鹵蟲有一定的致死活性, 致死率為20%。

化合物4: 白色無定形粉末,1H-NMR (CDCl3)δH: 6.09 (1H, s, H-3), 8.51 (1H, s, H-6), 6.07 (1H, d,J= 14.8Hz, H-7), 6.68 (1H, m, H-8), 1.94 (3H, d,J= 5.5 Hz, H-9), 7.08 (1H, s, H-3'), 6.64 (1H, s, H-5'), 3.86 (3H, s, H-8'), 3.78 (3H, s, H-9'), 3.78 (3H, s, H-10');13C NMR (CDCl3)δC: 161.4 (C-2, C), 115.0 (C-3, CH), 175.9 (C-4, C), 126.0 (C-5, C), 160.9 (C-6, CH), 122.7 (C-7, CH), 136.3 (C-8, CH), 18.6 (C-9, CH3), 191.7 (C-10, C), 126.1 (C-1', C), 129.9 (C-2', C), 105.3 (C-3', CH), 160.7 (C-4', C), 103.0 (C-5', CH), 157.4 (C-6', C), 166.4 (C-7', C), 55.6 (C-8', CH3), 56.1 (C-9', CH3), 52.4 (C-10', CH3)。其波譜數(shù)據(jù)與deoxyfunicone的文獻(xiàn)報(bào)道[13]一致, 該化合物為首次從P.pinophilum中分離得到。當(dāng)樣品濃度為0.1 mg/mL時(shí), 對鹵蟲有一定的致死活性, 致死率為35%。

化合物5: 無色晶體,1H-NMR (CDCl3)δH: 5.48 (1H, s, H-3), 5.60 (1H, s, H-6), 5.93 (1H, d,J= 15.0 Hz, H-7), 6.67 (1H, m, H-8), 1.91 (3H, d,J= 6.9 Hz, H-9), 6.87 (1H, s, H-3'), 6.54 (1H, s, H-5'), 3.86 (3H, s, H-8'), 3.76 (3H, s, H-9'), 3.86 (3H, s, H-10');13C NMR (CDCl3)δC: 161.4 (C-2, C), 104.1 (C-3, CH), 185.3 (C-4, C), 62.6 (C-5, C), 81.6 (C-6, CH), 124.1 (C-7, CH), 137.7 (C-8, CH), 18.6 (C-9, CH3), 190.8 (C-10, C), 119.8 (C-1', C), 134.2 (C-2', C), 101.5 (C-3', CH), 163.0 (C-4', C), 106.2 (C-5', CH), 159.1 (C-6', C), 167.6 (C-7', C), 55.8 (C-8', CH3), 56.1 (C-9', CH3), 52.9 (C-10', CH3)。其波譜數(shù)據(jù)與5, 6-epoxy- 3-deoxyfunicone的文獻(xiàn)報(bào)道[14]一致, 該化合物為首次從P.pinophilum中分離得到。當(dāng)樣品濃度為0.1 mg/mL時(shí),對鹵蟲有一定的致死活性, 致死率為39%。

化合物6: 黃色油狀物,1H NMR (DMSO-d6)δH: 6.55 (1H, s, H-2), 6.60 (1H, s, H-5), 2.05 (3H, s, H-7), 6.22 (1H, d,J= 2.0 Hz, H-2'), 6.25 (1H, d,J= 0.5 Hz, H-4'), 6.19 (1H, d,J= 0.8 Hz, H-6'), 3.69 (3H, s, H-7');13C NMR (DMSO-d6)δC: 132.2 (C-1, C), 116.7 (C-2, CH), 144.4 (C-3, C), 143.6 (C-4, C), 117.5 (C-5, CH), 124.7 (C-6, C), 19.3 (C-7, CH3), 142.9 (C-1', C), 108.8 (C-2', CH), 159.9 (C-3', C), 99.2 (C-4', CH), 158.0 (C-5', C), 105.7 (C-6', CH), 54.8 (C-7', CH3)。其波譜數(shù)據(jù)與5'-methoxy-6-methyl-biphenyl-3, 4, 3'-triol的文獻(xiàn)報(bào)道[15]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物7: 白色無定形粉末,1H NMR (DMSO-d6)δH: 6.42 (1H, s, H-2), 6.53 (1H, s, H-5), 1.85 (3H, s, H-7), 6.43 (1H, d,J= 2.4 Hz, H-4'), 6.10 (1H, d,J= 2.4 Hz, H-6'), 3.76 (3H, s, H-8');13C NMR (DMSO-d6)δC: 132.4 (C-1, C), 116.6 (C-2, CH), 145.0 (C-3, C), 143.9 (C-4, C), 115.9 (C-5, CH), 124.9 (C-6, C), 18.8 (C-7, CH3), 142.1 (C-1', C), 108.7 (C-2', C), 161.5 (C-3', C), 99.6 (C-4', CH), 162.0 (C-5', C), 108.9 (C-6', CH), 171.5 (C-7', C), 55.3 (C-8', CH3)。其波譜數(shù)據(jù)與altenusin的文獻(xiàn)報(bào)道[16]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物8: 黃色無定形粉末,1H-NMR (DMSO-d6)δH: 3.42 (1H, m, H-1), 2.92 (1H, d,J= 19.0, 6.4 Hz, Ha-2), 2.21 (1H, d,J= 19 Hz, Hb-2), 6.80 (1H, s, H-7), 6.74 (1H, s, H-9), 1.34 (3H, d,J= 6.9 Hz, H-10), 3.92 (3H, s, H-11);13C NMR (DMSO-d6)δC: 27.7 (C-1, CH), 42.4 (C-2, CH2), 195.6 (C-3, C), 147.5 (C-3a, C), 165.2 (C-5, C), 100.5 (C-5a, C), 163.8 (C-6, C), 103.9 (C-7, CH), 166.2 (C-8, C), 101.9 (C-9, CH), 134.7 (C-9a, C), 144.6 (C-10a, C), 20.5 (C-10, CH3), 56.1 (C-11, CH3)。其波譜數(shù)據(jù)與1-deoxyrubralactone的文獻(xiàn)報(bào)道[17]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物9: 白色無定形粉末,1H-NMR (DMSO-d6)δH: 6.33 (1H, s, H-3), 8.02 (1H, s, H-6), 4.29 (2H, s, H-7), 5.65 (1H, br s, 7-OH), 9.04 (1H, br s, 5-OH);13C NMR (DMSO-d6)δC: 168.0 (C-2, C), 109.8 (C-3, CH), 173.9 (C-4, C), 145.7 (C-5, C), 139.2 (C-6, CH), 59.4 (C-7, CH2)。其波譜數(shù)據(jù)與kojic acid的文獻(xiàn)報(bào)道[18]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物10: 黃色油狀物,1H-NMR (DMSO-d6)δH: 5.97 (1H, s, H-3), 6.61 (1H, s, H-6), 6.64 (1H, s, H-8), 2.58 (2H, m, H-1'), 4.17 (1H, m, H-2'), 1.13 (3H, d,J= 6.3 Hz, H-3'), 2.65 (3H, s, H-1'');13C NMR (DMSO-d6)δC: 164.4 (C-2, C), 111.4 (C-3, CH), 178.1 (C-4, C), 141.2 (C-5, C), 116.3 (C-6, CH), 160.9 (C-7, C), 100.3 (C-8, CH), 158.8 (C-9, C), 114.2 (C-10, C), 42.6 (C-1', CH2), 63.9 (C-2', CH), 23.3 (C-3', CH3), 21.9 (C-1'', CH3)。其波譜數(shù)據(jù)與7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone的文獻(xiàn)報(bào)道[19]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物11: 白色無定形粉末,1H NMR (DMSO-d6)δH: 6.36 (1H, s, H-4), 2.01～2.62 (6H, m, H-1, 2, 7), 2.81 (1H, m, H-9), 2.48 (2H, m, H-15), 1.69～2.02 (6H, m, H-11, 12, 16), 1.48 (1H, m, H-17), 0.98 (3H, s, H-18), 1.26 (3H, s, H-19), 2.42 (1H, m, H-20), 1.09 (3H, d,J= 6.8 Hz, H-21), 5.22 (1H, d,J= 15.3 Hz, H-22), 5.18 (1H, dd,J= 15.3, 7.7 Hz, H-23), 1.88 (1H, m, H-24), 1.47 (1H, d,J =6.8, H-25), 0.82 (3H, d,J= 7.2 Hz, H-26), 0.83 (3H, d,J= 7.2 Hz, H-27), 0.92 (3H, d,J= 7.2 Hz, H-28);13C NMR (DMSO-d6)δC: 38.9 (C-1, CH2), 34.2 (C-2, CH2), 198.8 (C-3, C), 124.5 (C-4, CH), 157.3 (C-5, C), 199.9 (C-6, C), 40.8 (C-7, CH2), 62.2 (C-8, C), 49.4 (C-9, CH), 36.0 (C-10, C), 25.1 (C-11, CH2), 38.3 (C-12, CH2), 53.9 (C-13, C), 215.1 (C-14, C), 37.9 (C-15, CH2), 23.2 (C-16, CH2), 49.4 (C-17, CH), 17.1 (C-18, CH3), 24.0 (C-19, CH3), 37.3 (C-20, CH), 23.6 (C-21, CH3), 132.8 (C-22, CH), 133.6 (C-23, CH), 43.2 (C-24, CH), 33.1 (C-25, CH), 20.0 (C-26, CH3), 19.7 (C-27, CH3), 17.6 (C-28, CH3)。其波譜數(shù)據(jù)與 dankasterone的文獻(xiàn)報(bào)道[20]一致, 該化合物為首次從P.pinophilum中分離得到, 并對鹵蟲有較好的致死活性, LD50值為39.2 μmol/L, 而陽性對照LD50值為92.1μmol/L。

化合物12: 白色無定形粉末,1H-NMR (DMSO-d6)δH: 6.42 (1H, d,J= 2.4 Hz, H-3), 6.27 (1H, dd,J= 8.5, 2.4 Hz, H-5), 7.44 (1H, d,J= 8.5 Hz, H-6), 1.99 (3H, s, H-8), 3.73 (3H, s, H-9), 8.86 (1H, s, NH);13C NMR (DMSO-d6)δC: 118.7(C-1, C), 151.6 (C-2, C), 99.2 (C-3, CH), 154.9 (C-4, C), 106.0 (C-5, CH), 124.4 (C-6, CH), 167.8 (C-7, C), 23.4 (C-8, CH3), 55.3 (C-9, CH3)。其波譜數(shù)據(jù)與4-hydroxy-2-methoxyacetanilide的文獻(xiàn)報(bào)道[21]一致, 該化合物為首次從P.pinophilum中分離得到。

化合物13: 無色油狀物,1H-NMR (DMSO-d6)δH: 8.57 (1H, dd,J= 8.3, 0.8 Hz, H-3), 7.47 (1H, td,J= 7.7, 1.0 Hz, H-4), 7.11 (1H, td,J= 7.7, 1.0 Hz, H-5), 7.74 (1H, dd,J= 7.7, 1.0 Hz, H-6), 4.10 (1H, q,J= 6.8Hz, H-9), 1.30 (3H, d,J= 6.8 Hz, H-10), 12.0 (1H, br s, NH), 6.09 (1H, br s, 9-OH), 8.56 (1H, br s, CONHa), 7.56 (1H, br s, CONHb);13C NMR (DMSO-d6)δC: 120.9 (C-1, C), 138.7 (C-2, C), 119.8 (C-3, CH), 131.7 (C-4, CH), 122.3 (C-5, CH), 128.4 (C-6, CH), 170.2 (C-7, C), 174.0 (C-8, C), 67.8 (C-9, CH), 20.8 (C-10, CH3)。其波譜數(shù)據(jù)與 N-(2-hydroxypropanoyl)-2-aminobenzoic acid amide的文獻(xiàn)報(bào)道[22]一致, 該化合物為首次從P.pinophilum中分離得到。

[1] 屈建航, 李寶珍, 袁紅莉. 沉積物中微生物資源的研究方法及其進(jìn)展[J]. 生態(tài)學(xué)報(bào), 2007, 27(6): 2636-2641.

[2] 劉璞, 王紅梅. 海洋沉積物微生物多樣性研究[J]. 安徽農(nóng)業(yè)科學(xué), 2009, 37(10): 4569-4571.

[3] 黃永中, 羅雄明, 王建華. 海洋沉積物來源鏈霉菌屬次生代謝產(chǎn)物及其生物活性研究進(jìn)展[J]. 天然產(chǎn)物研究與開發(fā), 2011, 23(4): 758-766.

[4] Blunt J W, Copp B R, Hu W P, et al.Marine natural products [J]. Nat Prod Rep, 2008, 25(1): 35-94.

[5] Blunt J W, Copp B R, Hu W P, et al.Marine natural products [J]. Nat Prod Rep, 2009, 26(2): 170-244.

[6] Blunt J W, Copp B R, Munro M H G, et al. Marine natural products [J]. Nat Prod Rep, 2010, 27(2): 165-237.

[7] Wu Q X, Crews M S, Draskovic M, et al.Azonazine, a novel dipeptide from a Hawaiian marine sedimentderived fungus,Aspergillus insulicola[J]. Org Lett, 2010, 12(20): 4458-4461.

[8] Li C S, An C Y, Li X M, et al. Triazole and dihydroimidazole alkaloids from the marine sediment-derived fungusPenicillium paneumSD-44 [J]. J Nat Prod, 2011, 74(5): 1331-1334.

[9] Zhao Y, Chen H, Shang Z, et al. SD118-Xanthocillin X (1), a novel marine agent extracted fromPenicillium commune, induces autophagy through the inhibition of the MEK/ ERK pathway [J]. Mar Drugs, 2012, 10(6): 1345-1359.

[10] Gerwick W H, Proteau P J, Nagle D G, et al. Structure of curacin A, a novel antimitotic, antiproliferative, and brine shrimp toxic natural product from the marine CyanobacteriumLyngbya majuscule[J]. J Org Chem, 1994, 59(6): 1243-1245.

[11] Xia X K, Liu F, She Z G, et al.1H and13C NMR assignments for 6-demethylvermistatin and two penicillide derivatives from the mangrove fungusGuignardiasp. (No. 4382) from the South China Sea [J]. Magn Reson Chem, 2008, 46(7): 693-696.

[12] Komai S I, Hosoe T, Itabashi T, et al. New vermistatin derivatives isolated fromPenicillium simplicissimum[J]. Heterocycles, 2005, 65(11): 2771-2776.

[13] Sassa T, Nukina M, Suzuki Y. Deoxyfunicone, a new γ-pyrone metabolite from a resorcylide-producing fungus (Penicilliumsp.) [J]. Agric Biol Chem, 1991, 55(9): 2415-2416.

[14] Komai S I, Hosoe T, Itabashi T, et al.A new funicone derivative isolated fromTalaromyces flavusIFM 52668 [J]. Mycotoxins, 2004, 54(1): 15-19.

[15] Wang Q X, Bao L, Yang X L, et al. Polyketides with antimicrobial activity from the solid culture of an endolichenic fungusUlocladiumsp. [J].Fitoterapia, 2012, 83(1): 209-214.

[16] Cota1 B B, Rosa L H, Caligiorne R B, et al. Altenusin, a biphenyl isolated from the endophytic fungusAlternariasp., inhibits trypanothione reductase fromTrypanosoma cruzi[J]. FEMS Microbiol Lett, 2008, 285(2): 177-182.

[17] Naganuma M, Nishida M, Kuramochi K, et al. 1-deoxyrubralactone, a novel specific inhibitor of families X and Y of eukaryotic DNA polymerases from a fungal strain derived from sea algae [J]. Bioorg Med Chem, 2008, 16(6): 2939-2944.

[18] Kingsbury C A, Cliffon M, Looker J H. Crobon-13 nuclear magnetic resonance spectra of kojic acid and other 4-pyrone derivatives [J]. J Org Chem, 1976, 41(16): 2777-2780.

[19] Xu J, Kjer J, Sendker J, et al.Chromones from the endophytic fungusPestalotiopsissp. isolated from the Chinese mangrove plantRhizophora mucronata[J]. J Nat Prod, 2009, 72(4): 662-665.

[20] Amgata T, Doi M, Tohgo M, et al.Dankasterone, a new class of cytotoxic steroid produced by aGymnascellaspecies from a marine sponge [J]. Chem Commun, 1999, 35(14): 1321-1322.

[21] Findlay J A, Li G Q, Miller J D, et al. Insect toxins from spruce endophytes[J]. Can J Chem, 2003, 81(4): 284-292.

[22] Dai M C, Tabacchi R, Saturnin C. Nitrogen-containing aromatic compound from the culture medium ofPenicillium chrysogenumthom. Chimia, 1993, 47(6): 226-229.

(本文編輯: 康亦兼)

Study of metabolites fromPenicillium pinophilumSD-272, a marine sediment-derived fungus

WANG Ming-hui1,2, LI Xiao-ming1, LI Chun-shun1, WANG Bin-gui1
(1. Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China; 2. University of Chinese Academy of Sciences, Beijing 100049, China)

Mar., 23, 2013

sediment;Penicillium pinophilum; secondary metabolite; brine shrimp lethality

In this paper, thirteen compounds were isolated from the zymotic fluid extract of the fungal strainPenicillium pinophilumSD-272by a combination of silica gel, Sephadex LH-20, and Lobar LiChroprep RP-18 column chromatography as well as the preparative thin layer chromatography. SD-272was isolated from sediment sample collected from the estuary of the Pearl River in South China Sea. The structures of these compounds were elucidated mainly based on the analysis of the UV, MS, 1D and 2D NMR as 4′-demethylvermistatin (1), vermistatin (2), penisimplicissin(3), deoxyfunicone (4), 5, 6-epoxy-3-deoxyfunicone (5), 5′-methoxy-6- methyl-biphenyl-3, 4, 3′-triol (6), altenusin (7), 1-deoxyrubralactone(8), kojic acid (9), 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (10), dankasterone (11), 4-hydroxy-2-methoxyacetanilide (12), and N-(2-hydroxypropanoyl)- 2-aminobenzoic acid amide (13). All the compounds were firstly reported to be isolated fromP.pinophilum. Compound 11 displayed potent brine shrimp lethality with a LD50of 39.2 μmol/L.

O629

A

1000-3096(2014)03-0001-05

10.11759/hykx20130323002

2013-03-23;

2013-05-26

國家自然科學(xué)基金項(xiàng)目(31270403); 科技部“973計(jì)劃”項(xiàng)目(2010CB833802)

王鳴慧(1984-), 女, 內(nèi)蒙古錫盟人, 博士研究生, 主要研究方向?yàn)樘烊划a(chǎn)物化學(xué), E-mail: wangmh29@yahoo.cn; 王斌貴, 男,

, 研究員, 博士生導(dǎo)師, 電話: 0532-82898553, E-mail: wangbg@qdio.ac.cn