1.1.1.346 F22Y the mutation causes a 2.5fold decrease in Km for 2,5-didehydro-D-gluconate whereas the value of kcat remains essentially unchanged 440307 1.1.1.346 F22Y/A272G substrate-binding pocket double mutant with decreased kcat value for NADPH compared to the wild type enzyme 440311 1.1.1.346 F22Y/K232G/R235G/R238H/A272G mutant with wild type kcat value for NADPH 440311 1.1.1.346 F22Y/K232G/R235T/R238H/A272G 420 mutant with decreased kcat value for NADPH compared to the wild type enzyme 440311 1.1.1.346 F22Y/K232G/R238H/A272G mutant with decreased kcat value for NADPH compared to the wild type enzyme 440311 1.1.1.346 F22Y/K232G/R238H/A272G the mutant exhibits activity with NADH that is more than 2 orders of magnitude higher than that of the wild type enzyme and retains a high level of activity with NADPH 440311 1.1.1.346 F22Y/K232G/R238H/A272G the mutation enhances binding to NADH, while retaining to a large extent the ability to bind NADPH. The mutant is also more stable and can, therefore, be expected to exhibit greater effective activity at elevated temperatures in comparison to the wild type enzyme 657308 1.1.1.346 K232G/R238H mutant with decreased kcat value for NADPH compared to the wild type enzyme 440311 1.1.1.346 K233G the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme 440310 1.1.1.346 K233H the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 K233M the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme 440310 1.1.1.346 K233Q the mutant shows wild type NADPH activity and increased NADH activity 440310 1.1.1.346 K233R the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 K233S the mutant shows wild type NADPH activity and increased NADH activity compared to the wild type enzyme 440310 1.1.1.346 K233T the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 additional information construction of enzyme gene knockout mutant M-AKR, that shows decreased degradation activity with testosterone, estradiol, oestrone, and methyltestosterone compared to the wild-type enzyme. Compared to the wild-type, the mutation of the endogenous 2,5DKR gene results in lower degradation of estradiol and methyltestosterone but has no effct on degradation of estrone and testosterone -, 742259 1.1.1.346 additional information establishment of an efficient process for 2,5-DKG reductase production that also satisfies food safety requirements. Food grade variants of the Lactobacillales based expression systems pSIP (Lactobacillus plantarum) and NICE (Lactococcus lactis) are evaluated with regard to their effictiveness to produce 2,5-DKG reductase from Corynebacterium glutamicum, overview. Lactobacillus plantarum/pSIP609 is an interesting alternative to Escherichia coli expression systems for industrial 2,5-DKG reductase production. Highest production levels of 2,5-DKG reductase are obtained with the system Lactobacillus plantarum/pSIP609, resulting in 104 U/l without pH regulation and 262 U/l with pH control at pH 6.5 -, 741575 1.1.1.346 Q192R the mutation primarily affects the kcat parameter toward the 2,5-didehydro-D-gluconate substrate, increasing its value approximately 2.5fold, whereas Km is relatively unaffected, or increases slightly 440307 1.1.1.346 R235C the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235D the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 R235E the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 R235G the mutant shows decreased NADPH activity and increased NADH activity compared to the wild type enzyme 440310 1.1.1.346 R235H the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235M the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235N the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235Q the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235S the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R235T the mutant shows wild type NADPH activity and increased NADH activity 440310 1.1.1.346 R235Y the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 R238D the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 R238E the mutant shows no activity with NADPH and increased NADH activity compared to the wild type enzyme 440310 1.1.1.346 R238F the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 R238G the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 R238H the mutant shows wild type NADPH activity and increased NADH activity 440310 1.1.1.346 R238N the mutant shows reduced NADPH activity and no NADH activity 440310 1.1.1.346 R238Q the mutant shows reduced NADPH activity compared to the wild type enzyme and no NADH activity 440310 1.1.1.346 R238Y the mutant shows reduced NADPH activity and increased NADH activity ompared to the wild type enzyme 440310 1.1.1.346 S233E the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 S233K the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 S233M the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 S233N the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 S233T the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 S233V the mutant shows no activity with NADPH and NADH 440310 1.1.1.346 V234D the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234E the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234I the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234M the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234M/R235C the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234N the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234Q the mutant shows wild type NADPH activity and no NADH activity 440310 1.1.1.346 V234S the mutant shows decreased NADPH activity compared to the wild type enzyme and no NADH activity 440310