化学诱变结合结构类似物法选育高苯丙氨酸解氨酶菌种

ＥＭＳ诱变结合结构类似物法提高含ＰＡＬ出发菌株红酵母Ｒｈｏｄｏｔｏｒｕｌａ ｓｐ．ＣＩＢＡＳＡ１４０１１００ 生物转化能力阳性率为３５．５ ％ ，部分突变株ＰＡＬ转化能力提高１ 倍以上，其中Ｅ１０５，Ｅ２４０ 具有快速转化能力，反应３ ｈＬＰｈｅ累积浓度达到１ ｇＬ左右．φ１０％ 甘油和３０ ｇＬＧｌｕ 对ＰＡＬ转化具有明显的稳定作用．用含φ１０％ 甘油和３０ ｇＬＧｌｕ 的反应液进行生物转化制备，Ｅ１０５ 和Ｅ２４０ 在２４ ｈ 期间ＬＰｈｅ 质量浓度大于２０ ｇＬ，转化率达８０ ％ ．

BREEDING FOR HIGH L-PHENYLALANINE AMMONIA-LYASE CONTAINING STRAINS BY MEANS OF CHEMICAL MUTAGENESIS COMBINED WITH STRUCTURE ANALOGS

The original strain Rhodotorula sp.CIBAS A 1401 100 containing phenylalanine ammonia lyase(PAL) was bred by means of EMS mutagenesis in combination with structure analogs. Among the mutants investigated, the positive ratio of PAL activity improvement was 35.5%. Some mutants showed twice higher activity than the parent. Mutants E105 and E240 had fast bioconversion capacity and they produced L Phe 1 g/L within 3 h. 10% glycerol and 30 g/L glutamate were very effective for the stabilization of PAL bioconversion. In the preparative-scale bioconversion using 10% glycerol and 30 g/L glutamate as stabilizers, E105 and E240 made the L Phe yield over 20 g/L and conversion ratio over 80% during 24 h.