不同抗生素和防腐剂对小球藻Chlorella sorokiniana细胞生长的影响

Effects of different antibiotics and preservatives on Chlorella sorokiniana cell growth

  • 摘要: 目的分析不同抗生素和防腐剂对小球藻Chlorella sorokiniana细胞生长的影响,筛选出适宜的抗生素和防腐剂种类及其剂量,为建立小球藻C.sorokiniana无菌培养体系提供参考依据.方法在HSM培养基中分别添加10、20、40、60、80和100μg/mL抗生素,包括氯霉素、链霉素、青霉素、庆大霉素和头孢噻肟,以及山梨酸钾(50、100、150和200μg/mL)、乳酸钠(0.5%、1.0%和2.0%)、富马酸二甲酯(0.1、0.5、1.0、5.0、10.0、25.0和50.0μg/mL)等防腐剂,培养3 d后测定小球藻C.sorokiniana的细胞密度、叶绿素含量及光系统Ⅱ最大光化学量子产量(Fv/Fm).结果添加10~100μg/mL青霉素或10~80μg/mL头孢噻肟对小球藻C.sorokiniana细胞密度、叶绿素含量和Fv/Fm的影响均不显著(P>0.05,下同);添加100μg/mL头孢噻肟会抑制藻细胞生长,抑制率为18.89%,并引起叶绿素含量和Fv/Fm极显著下降(P<0.01,下同).氯霉素、链霉素和庆大霉素等蛋白质合成抑制类抗生素对小球藻C.sorokiniana细胞的毒性较强,高于10μg/mL的链霉素和庆大霉素及高于20μg/mL的氯霉素均极显著抑制藻细胞生长,对应的叶绿素含量和Fv/Fm也极显著下降.3种防腐剂中,山梨酸钾对小球藻C.sorokiniana细胞生长无显著影响,添加200μg/mL山梨酸钾还会促进细胞叶绿素含量增加;乳酸钠对小球藻C.sorokiniana细胞有一定的毒性作用,其藻细胞密度、叶绿素含量和Fv/Fm随添加浓度增加而降低;添加0.1~10.0μg/mL富马酸二甲酯不影响小球藻C.sorokiniana的细胞密度及叶绿素含量,但Fv/Fm极显著下降.结论建立小球藻C.sorokiniana无菌培养体系时宜选用青霉素和头孢噻肟为抗生素、山梨酸钾为防腐剂,其推荐使用浓度分别为青霉素100μg/mL、头孢噻肟80μg/mL、山梨酸钾200μg/mL.

     

    Abstract: ObjectiveEffects of different antibiotics and preservatives on growth of Chlorella sorokiniana cells were analyzed,and optimal antibiotics and preservatives and doses were screened to provide reference for establishing axenic culture of C. sorokiniana.MethodAntibiotics and preservatives were added in HSM media including chloramphenicol, streptomycin,penicillin,gentamicin and cefotaxime sodium(10,20,40,60,80 and 100μg/mL),potassium sorbate (50,100,150 and 200μg/mL),lactic acid bacteria(0.5%,1.0%and 2.0%),dimethyl fumarate(0.1,0.5,1.0,5.0, 10.0,25.0 and 50.0 μg/mL). Cell density,chlorophyll content and photosystem Ⅱ maximal photochemical efficiency (Fv/Fm)were determinated after 3 d cultivation.ResultCell density,chlorophyll content and Fv/Fm of C. sorokiniana were not significantly affected by 10-100 μg/mL penicillin and 10-80 μg/mL cefotaxime sodium(P>0.05 ,the same below). Adding 100μg/mL cefotaxime sodium inhibited cell growth with inhibition rate of 18.89%,and also decreased chlorophyll content and Fv/Fm extremely significantly(P<0.01,the same below). Compared with control,protein synthesis inhibitory antibiotics like chloramphenicol ,streptomycin and gentamicin were more toxic to C. sorokiniana. Strepto-mycin and gentamicin with concentrations over 10 μg/mL and chloramphenicol with concentrations over 20μg/mL had extremely significant inhibition effects on cell growth ,and the corresponding chlorophyll content and Fv/Fm dropped extremely significantly. Among the three preservatives,potassium sorbate did not affect C. sorokiniana cell growth sig-nificantly. Furthermore,the chlorophyll content was enhanced by 200μg/mL potassium sorbate. Lactic acid bacteria was toxic to C. sorokiniana cell at certain extent,cell density,chlorophyll content and Fv/Fm of C. sorokiniana decreased as the concentration increased. Adding 0.1-10.0 μg/mL dimethyl fumarate did not affect the cell density and chlorophyll content of C. sorokiniana,but the Fv/Fm significantly decreased.ConclusionIn establishing axenic culture of C. soroki-niana ,the suitable antibiotics are penicillin and cefotaxime sodium ,suitable preservative is potassium sorbate. The optimal concentration are 100μg/mL(penicillin),80μg/mL(cefotaxime sodium)and 200μg/mL(potassium sorbate) respectively.

     

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