Transaminase mutant and application thereof
A transaminase and mutant technology, applied in the field of transaminase mutants and their applications, can solve the problem that transaminase is difficult to catalyze the synthesis of large-sterically hindered chiral amine compounds
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Embodiment 1
[0076] Directed mutations were carried out on the basis of the mother parent. The specific mutation sites are shown in the table below, and the catalytic activity of the mutants was tested according to the following reaction conditions:
[0077] 1 mL system includes 1 mg substrate 1, 1 mg PLP, 1 mg isopropylamine hydrochloride, 50 mg enzyme powder, pH8.0 100mM phosphate buffer, react for 40 h.
[0078] The test results are shown in the table below:
[0079] Table 1:
[0080]
[0081] .
[0082] Note: In the above table, 0 means the conversion rate is <1%, + means the conversion rate is greater than or equal to 1% and less than 5%, and ++ means the conversion rate is greater than or equal to 5% and less than or equal to 10%.
Embodiment 2
[0084] On the basis of Example 1, the combined mutation was carried out, and the activity of the combined mutation was screened according to the same reaction conditions as in Example 1. The results are shown in the table below.
[0085] Table 2:
[0086]
[0087] .
[0088] Note: In the above table, 0 means the conversion rate is <1%, + means the conversion rate is greater than or equal to 1% and less than 5%, ++ means the conversion rate is greater than or equal to 5% and less than 10%, +++ means the conversion rate is greater than or equal to 10 % and less than 15%, ++++ means the conversion rate is greater than or equal to 15% and less than 20%.
Embodiment 3
[0090] On the basis of Example 2, combined mutations were carried out, and the activity of combined mutations was screened according to the same reaction conditions as in Example 1. The results are shown in the table below.
[0091] table 3:
[0092]
[0093] .
[0094] Note: 0 means the conversion rate is <1%, + means the conversion rate is greater than or equal to 1% and less than 5%, ++ means the conversion rate is greater than or equal to 5% and less than 10%, +++ means the conversion rate is greater than or equal to 10% and less than 15% %, ++++ means the conversion rate is greater than or equal to 15% and less than 20%, and +++++ means the conversion rate is greater than or equal to 20% and less than or equal to 40%.
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