CAJ | 학술논문

从大西洋深海沉积物中分离得到1株产黑色素细菌bIp-2，16S rRNA基因序列分析表明该菌隶属于假单胞菌属（Pseudomonas），与施氏假单胞菌（P．stutzeri）有最高的16S rRNA基因序列相似性（99％）．进一步对该菌的生长及产黑色素条件进行了初步研究，结果表明bIp-2在稳定生长期后期才开始大量积累黑色素；bIp-2可在4～37℃的温度范围下生长，最适生长温度为37℃，在28℃具有最高的黑色素产量；bIp-2生长的pH范围偏碱性，在pH＝7的最适生长条件下黑色素产量最高，同时碱性环境有利于黑色素的快速形成；bIp-2可以在1％～9％的NaCl浓度范围中生长，在5％的NaCl浓度下具有最高的黑色素产量；低浓度的 Fe2＋（0．05 mmol/dm3）可以提高黑色素的产量， Fe2＋浓度≥0．20 mmol/dm3时明显抑制黑色素的生成，而Fe2＋浓度为1．00 mmol/dm3时可完全抑制菌株的生长；低浓度的 Cu2＋（0．1 mmol/dm3）对黑色素的产量无明显影响，Cu2＋浓度≥0．5 mmol/dm3时可抑制黑色素的生成，而Cu2＋浓度超过1．0 mmol/dm3时可完全抑制菌株的生长．通过PCR扩增获得菌株bIp-2的对羟苯基丙酮酸双加氧酶（hydroxyphenylpyruvic acid dioxygenase， HPPD）全长基因（1087 bp ），分析发现其编码的 HPPD 蛋白序列（含361氨基酸）与来自菌株Pseudomonas stutzeri RCH2的HPPD蛋白序列具有最高的同源性（序列相似性为97％）．这些结果为深入研究该细菌所产黑色素的合成，应用及生态作用奠定了基础．
종대서양심해침적물중분리득도1주산흑색소세균bIp-2，16S rRNA기인서렬분석표명해균대속우가단포균속（Pseudomonas），여시씨가단포균（P．stutzeri）유최고적16S rRNA기인서렬상사성（99％）．진일보대해균적생장급산흑색소조건진행료초보연구，결과표명bIp-2재은정생장기후기재개시대량적루흑색소；bIp-2가재4～37℃적온도범위하생장，최괄생장온도위37℃，재28℃구유최고적흑색소산량；bIp-2생장적pH범위편감성，재pH＝7적최괄생장조건하흑색소산량최고，동시감성배경유리우흑색소적쾌속형성；bIp-2가이재1％～9％적NaCl농도범위중생장，재5％적NaCl농도하구유최고적흑색소산량；저농도적 Fe2＋（0．05 mmol/dm3）가이제고흑색소적산량， Fe2＋농도≥0．20 mmol/dm3시명현억제흑색소적생성，이Fe2＋농도위1．00 mmol/dm3시가완전억제균주적생장；저농도적 Cu2＋（0．1 mmol/dm3）대흑색소적산량무명현영향，Cu2＋농도≥0．5 mmol/dm3시가억제흑색소적생성，이Cu2＋농도초과1．0 mmol/dm3시가완전억제균주적생장．통과PCR확증획득균주bIp-2적대간분기병동산쌍가양매（hydroxyphenylpyruvic acid dioxygenase， HPPD）전장기인（1087 bp ），분석발현기편마적 HPPD 단백서렬（함361안기산）여래자균주Pseudomonas stutzeri RCH2적HPPD단백서렬구유최고적동원성（서렬상사성위97％）．저사결과위심입연구해세균소산흑색소적합성，응용급생태작용전정료기출．
A melanin-producing strain was isolated from deep-sea sediment of the Atlantic Ocean and identified as Pseudomonas sp.bIp-2 by 16S rRNA gene sequencing.The study of growth and melanin production with varied culture conditions show that the melanin production in strain bIp-2 is growth-associated.The maximum yield of pig-ment was obtained after the culture attained the stationary growth phase.The temperature range for growth is 4~37℃ with optimal growth at 37℃,and the maximum melanin production was observed at 28℃.The pH range for growth is 6.0~9.0,with both optimal growth and melanin formation both being at pH 7.0.Strain bIp-2 grew in 1%~9%NaCl with a maximum melanin production at 5%NaCl.Low concentration of Fe(II)(0.05 mmol/dm3 ) can improve the production of melanin.Fe2+of more than 0.20 mmol/dm3 suppressed the melanin production and Fe2+of more than 1 .00 mmol/dm3 completely inhibited the growth of the strain.Cu2+of 0.1 mmol/dm3 had no effect to the melanin production and Cu2+of more than 0.5 mmol/dm3 inhibited the formation of melanin.Cu2+over 1 .0 mmol/dm3 completely inhibited the growth of the strain.A 1 087 bp fragment of the full-length hppD gene was amplified from the strain bIp-2.The hppD gene encoded an amino acid sequence sharing a 97%similarity with the HPPD from Pseudomonas stutzeri RCH2.These results laid a foundation to the study of the synthesis,applica-tion and ecological effect of the melanin from Pseudomonas sp.bIp-2.