砂土法、矿油封藏法保存链霉菌17年效果的评定

AN EVALUATION OF THE EFFICACY OF SAND-SOIL ADSORPTION AND MINERAL OIL SEAL METHOD FOR PRESERVATION OF STREPTOMYCES CULTURES FOR 17 YEARS

本文总结了用砂土法保存130个种、14个变种、409株链霉菌以及矿油封藏法保存其中103个种、11个变种、307株菌17年的效果;其中部分菌株的抗菌素产生能力和蛋白酶活力的测定结果;经矿油封藏后,死亡菌株的矿油层,用红外光谱法检查的结果。用砂土法保存的409株菌,17年后存活384株,存活率为93.9%;在144种（包括变种）中,有101种（70%）保持着原来的形态特征。用矿油封藏法保存的307株菌,17年后存活256株,存活率为83.4%;其中169株（66%）保持着原来的形态特征。产生链霉素的灰色链霉菌（Streptomyces griscus）As 4.181、比基尼链霉菌（S.bikiniensis）AS 4.569,产氯霉素的委内瑞拉链霉菌（S.venezu（?）la（?））AS 4.223等菌株,用矿油封藏法比砂土法保存得到较好的效果。产生新霉素的弗氏链霉菌（S.fradiae）AS 4.576,用砂土法保存效果较好。矿油封藏法保存链霉菌对保持蛋白酶活力,有着良好的效果。经红外光谱法检查19株失活的菌株的矿油层,其中17个样品于1710厘米-1或1600厘米-1处有吸收带。根据红外光谱,此吸收带属于脂肪族或芳香族中（?）c=c（?）基的伸展振动,与 Arai 用此法保存诺卡氏菌,分枝杆菌测出的结果近似。故推测链霉菌经此法保存后,菌株死亡的原因是由于浸出了细胞中类脂所致。砂土法和矿油封藏法对于长期大量保存链霉菌,是两种简便而且可取的方法。

In the present investigation theefficacy of sand-soil （2:1） adsorptionmethod,and mineral oil seal method forthe preservation of 130 species,14 varie-ties,and 409 strains of Streptomycesthrough a period of 17 years has beenanalyzed and compared,including theabilities of antibiosis and the activities ofproteinase;and the absorption spectra ofminerral oil layers of 19 dead cultureskept in paraffin oil by infrared spectro-photometry.Viability tests of these cultures areas follows:Of the 409 cultures preservedin sand-soil for 17 years,384 （93.9%）were viable;among 144 species （includ-ing the varieties）,101 species （70%） re-tained their originally morphological andcultural characteristics.Of the 307 cul-tures preserved under paraffin oil,256（83.4%） were viable;among the viablestrains,169 （66%） retained their inher-ent properties.In comparison with sand-soil adsorp-tion method or periodic transfer method,mineral oil seal for preservation of theabilities of Streptomyces griseus AS 4.181and S.bikiniensis AS 4.569 in producingstreptomycin,S.venezuelae AS 4.223 inproducing chloamphenicol were shown tohave better results.In general,the mineral oil seal me-thod for maintaining the activities of pro-teinase in some Streptomyces cultures ex-hibits good results.Infrared spectra of mineral oil layersanalyzed of 19 dead cultures,17 samplesshowed strong absorption bands near 1710and 1600cm-1.According to infraredspectroscopy,the bands at 1710 and 1600cm-1 indicate the stretching vibrations ofC=O and （?） C=C （?） of aliphatic com-pounds,respectively.Arai and his col-laborator have observed that some strainsof Nocardia and Mycobacterium werefound to be unable to survive for longstorage periods when sealed under paraffinoil;and that the paraffin oil layersshowed strong absorption bands near 1740and 1710 cm-1 when analyzed by infraredspectrophotometry.They suggested thatthe reasons for poor longevity of Nocardiaand Mycobacterium by oil seal preserva-tion might be ascribed to the removal ofcellular lipid components into oil layer.The results of the present investigationare similar to theirs.It is only differentfrom theirs in the band at 1740cm-1.However,the absorption spectrum of C=Oband has a wide wave number regionfrom 1600—1900cm-1 in different car-boxylic acids.The reasons for death ofsome Streptomyces cultures preservedunder mineral oil were ascribed to the re-moval of cellular lipid components into oillayer,because Streptomyces,Nocardia, and Mycobacterium are all prokaryotes,and are taxonomically related.Based on the results mentioned above,sand-soil adsorption method for keepingthe morphological characteristics of Strep-（?）omyces cultures can have better results.Although the viability proportion of themineral oil sealed cultures was lower thanthat of sand-soil adsorption method,theformer had better result for retainingthe physiological properties of someStreptomyces cultures.These two methodsbeing all simple,and not requiring specialequipment,and can therefore be simul-taneouly used to preserve Streptomycescultures on a large scale.