CAJ | 학술논문

2006～2007年从新疆罗布泊外嗣地区的泥土样品中分离了117株放线菌,初步筛选后对其中47株嗜碱放线菌进行了形态学鉴定,不同生长条件(pH值,盐NaCl、KCl、MgCl_2,碱性物质Na_2 CO_3、K_2CO_3、Mg_2CO_3、NaOH、KOH,温度)对菌株生长的影响,拮抗性、产酶活性检测及菌株LA4的分子进化鉴定等研究.结果发现,47株放线菌的15%属于诺卡氏菌属,70%属于链霉菌属,10%属于游动放线菌属,其中嗜碱链霉菌属占优势.在pH 7～11范围内,47株嗜碱放线菌均能生长.生长率为92%～95%.NaCl、KCl、MgCl,对嗜碱放线菌均具有抑制作用,NaCl、KCl的抑制作用高于MgCl_2,NaCl、KCl浓度超过10%时而MgCl_2 浓度超过15%时嗜碱放线菌完全停止生长.47株嗜碱放线菌对Na_2CO_3、K_2CO_3、MgCO_3、KOH、NaOH等碱性物质没有选择性,生长良好.47株嗜碱放线菌的最适生长温度为28～30℃.超过55℃后均不能生长.从47株嗜碱放线菌中筛选出了30株具有抗菌活性的菌株,其中对大肠杆菌有抑制作用的有11株,占36.7%,对金黄色葡萄球菌有抑制作用的有20株,占66.7%,对枯草杆菌有抑制作用的有16株,占53.3%;其中4株嗜碱放线菌LA4、LA24、LA33和LA19对3种指示菌均表现出抑制作用.在47株嗜碱放线菌中,74%具有淀粉酶活性,81%具有纤维素酶活性,43%具有蛋白酶活性,49%具有脂肪酶活性.通过分子鉴定,发现LA4为链霉菌中尚未报道的一个新亚种,其16S序列GenBank登录号为FJ182229.本研究结果表明新疆罗布泊外围地区存在大量的产纤维素酶、淀粉酶的嗜碱放线菌,并且潜藏着新的放线菌资源,为该地区嗜碱放线菌的种质资源开发和新药筛选有效菌源提供了参考.图5表4参20
2006～2007년종신강라포박외사지구적니토양품중분리료117주방선균,초보사선후대기중47주기감방선균진행료형태학감정,불동생장조건(pH치,염NaCl、KCl、MgCl_2,감성물질Na_2 CO_3、K_2CO_3、Mg_2CO_3、NaOH、KOH,온도)대균주생장적영향,길항성、산매활성검측급균주LA4적분자진화감정등연구.결과발현,47주방선균적15%속우낙잡씨균속,70%속우련매균속,10%속우유동방선균속,기중기감련매균속점우세.재pH 7～11범위내,47주기감방선균균능생장.생장솔위92%～95%.NaCl、KCl、MgCl,대기감방선균균구유억제작용,NaCl、KCl적억제작용고우MgCl_2,NaCl、KCl농도초과10%시이MgCl_2 농도초과15%시기감방선균완전정지생장.47주기감방선균대Na_2CO_3、K_2CO_3、MgCO_3、KOH、NaOH등감성물질몰유선택성,생장량호.47주기감방선균적최괄생장온도위28～30℃.초과55℃후균불능생장.종47주기감방선균중사선출료30주구유항균활성적균주,기중대대장간균유억제작용적유11주,점36.7%,대금황색포도구균유억제작용적유20주,점66.7%,대고초간균유억제작용적유16주,점53.3%;기중4주기감방선균LA4、LA24、LA33화LA19대3충지시균균표현출억제작용.재47주기감방선균중,74%구유정분매활성,81%구유섬유소매활성,43%구유단백매활성,49%구유지방매활성.통과분자감정,발현LA4위련매균중상미보도적일개신아충,기16S서렬GenBank등록호위FJ182229.본연구결과표명신강라포박외위지구존재대량적산섬유소매、정분매적기감방선균,병차잠장착신적방선균자원,위해지구기감방선균적충질자원개발화신약사선유효균원제공료삼고.도5표4삼20
117 stains of actinomycetes were isolated from the soil and mud samples collected from the peripheral areas of the Lop Nur in Xinjiang in 2006 and 2007. Of them, 47 strains were alkalophilic actinomycetes. Their morphological characters were analyzed, and their growth features (pH; NaCl, KC1 and MgCl_2; alkaline substances Na_2CO_3, K_2CO_3, MgCO_3, NaOH and KOH; temperature), antagonistic actions and enzyme activities, as well as the molecular level of strain LA4 were studied. The results showed that 15% of the 47 strains were initially identified belonging to Nocardia, 70% to Streptomyces, 10% to Actinoplanes; and the 47 strains could grow at pH range of 7～11, and their growth rate ranging between 92% and 95%. NaCl, KC1 and MgCl_2 could inhibit the growth of all the alkalophilic actinomycetes. The inhibitory effect of NaCl and KC1 was higher than that of MgCl_2. When the concentrations of NaCl and KC1 exceeded 10%, and that of MgCl_2 exceeded 15%, the alkalophilic actinomycetes stopped growing. The alkalophilic actinomycetes could grow well on alkaline substances of Na_2CO_3, K_2CO_3, MgCO_3, NaOH and KOH, with no selection of them. Their optimal growth temperature was 28~30 ℃. When the temperature was over 55 ℃, the alkalophilic actinomycetes could not grow. Of them, 30 strains with antibacterial activity were selected, and 11 of them had antagonistic action on Escherichia coli, accounting for 36.7%, 20 of them on Staphylococcus aureus, for 66.8%, 16 of them on Bacillus subtilis, for 53.3% and 4 of them LA4, LA24, LA33 and LA19 on the three kinds of indicator bacteria. 74% of the 47 alkalophilic actinomycetes produced amylase, 81% produced cellulase, 43% produced protease and 49% produced lipase. Based on 16S sequence, LA4 was identified as a new subspecies of Streptomyces. The sequence of LA4 was obtained from the GenBank with its number of FJ18229. This study indicated that the peripheral area of Lop Nur was found with large number actinomycetes as new resources for producing amylase and cellulase. Fig 5, Tab 4, Ref 19