摘要
对生淀粉糖化菌黑曲霉Sx用混和酶制取原生质体进行了探索.在GM培养基中加入一定量的Cu2+、Mn2+,原生质体产量较高;采用pH4~5的蜗牛酶、纤维素酶、溶菌酶的混合酶,最佳浓度比为:1.5∶3.5∶1.5(mg/mL).取在GM培养基中培养17~18h的菌丝,在32℃酶解3h,原生质体产量最高,达到2.4×105个/mL;在加入β-巯基乙醇及二硫苏糖醇处理菌丝时,原生质体产量提高近3倍.在再生培养基中,加入终浓度为30mmol/LMgSO4时,原生质体再生率由16.7%提高到25.6%.用He-Ne激光照射原生质体时,功率9mW照射30min时致死率为50%,随着时间的延长,原生质体存活率降低.采用波长632.5nm、功率9mW、光斑直径5mm的He-Ne激光多次照射诱变,筛选出一株酶活力最高菌株黑曲霉Sy,活力提高51%,经连续传代5次。
This article deals with the formation and regeneration of the protoplast
from A.niger Sx.The protoplast production is relatively high when Cu 2+ or Mn 2+ was
added to the growth media.2.4×10 5/mL protoplasts can be obtained at 32℃ lasting 3 hours
from 17~18 hours cell age mycelia,using the mixture of Snailase (1.5mg/mL),Cellulase
(3.5mg/mL) and Lysozyme (1.5mg/mL) with pH 4~5.Treated with β Mercaptoethanol and
DTT,the production of protoplast was nearly three times of the original one.When the
concentration of MgSO 4 in RM was 30mmol/L,the regeneration rate increased from 16.7% to
25.6%.With the He Ne laser (λ have 632.8nm,the power have 9mW,the diameter have Φ5mm)
irradiating 30 minutes,the death rate of protoplast is 50%,and the He Ne is used to mutagenize
the protoplast,through a large amount of screening,a high activity raw starch
amylomyces,named A.niger Sy.The enzyme activity is increased by 51%.
出处
《光子学报》
EI
CAS
CSCD
1999年第7期603-606,共4页
Acta Photonica Sinica
基金
陕西省科学技术委员会自然科学基金
关键词
黑曲霉
选育
生淀粉糖化菌
原生质体
激光诱变
Aspergillus niger
He Ne laser
Selective breeding
Raw starch amylomyces
Protoplast