Transaminase mutant and application thereof
A technology of transaminase and mutant, which is applied to the mutant of transaminase and its application field, and can solve the problem that transaminase is difficult to catalyze synthesis of large sterically hindered chiral amine compounds and the like
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Embodiment 1
[0084] The directional mutation was carried out on the basis of the parent parent. The specific mutation sites are shown in the following table, and the catalytic activity of the mutant was tested according to the following reaction conditions:
[0085] 1 mL system includes 1 mg substrate 1, 1 mg PLP, 1 mg isopropylamine hydrochloride, 50 mg enzyme powder, pH 8.0 100 mM phosphate buffer, 50 o C reacted for 40 h.
[0086] The test results are shown in the table below:
[0087] Table 1:
[0088]
[0089] .
[0090] Note: In the above table, 0 means the conversion rate is less than 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
[0092] On the basis of Example 1, combined mutation was performed, and the activity of the combined mutation was screened according to the same reaction conditions as Example 1. The results are shown in the following table.
[0093] Table 2:
[0094]
[0095] .
[0096] Note: In the above table, 0 means the conversion rate is less than 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%, and +++ 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
[0098] On the basis of Example 2, combined mutation was carried out, and the activity of the combined mutation was screened according to the same reaction conditions as Example 1. The results are shown in the following table.
[0099] table 3:
[0100]
[0101] .
[0102] Note: 0 means conversion rate <1%, + means conversion rate is greater than or equal to 1% and less than 5%, ++ means conversion rate is greater than or equal to 5% and less than 10%, +++ means 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%, +++++ means the conversion rate is greater than or equal to 20% and less than or equal to 40%.
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