Table 1 Advancements in enzyme engineering for functional lipids synthesis

Activity

Phospholipase D (PLD)

For the enzymatic production of phosphatidylserine

Directed evolution

The mutation demonstrating a 3.24-fold increase in transphosphatidylation conversion compared to the WT

(Zhang et al. 2019a)

PLD

For the enzymatic production of phosphatidylserine

Substrate pocket reconstruction strategy

The mutant displayed 2.04-fold increase in the transphosphatidylation/hydrolysis ratio compared to the WT

(Qi et al. 2022)

Candida antarctica lipase A (CALA)

For the enrichment of long chain mono-unsaturated fatty acids

Reshaping of binding tunnels

The variant V290W doubled C20:1 in the esterified fraction from 15 to 34%

(Zorn et al. 2019, 2018)

Selectivity

Lipase MAS1

For the enzymatic production of DAGs

Substrate binding pocket engineering

The mutation showed an increased synthesis ratio of partial glycerides/triglycerides to 6.32, compared to 1.21 in the WT

(Yang et al. 2022)

Candida antarctica lipase B (CALB)

For the enzymatic production of 1-monoacyl-sn-glycerol

Substrate binding pocket engineering

The mutation showed twofold increase in selectivity for synthesizing 1-monoacyl-sn-glycerol

(Woo et al. 2022)

PLD

For PLD selectivity, the positional specificity toward the 1-OH of myo-inositol

Substrate binding pocket engineering

The mutation showed remarkable 98% positional specificity

(Samantha et al. 2021)

Fatty acid hydratases

For the enzymatic production of high-value HFAs

Sequence alignment and structure analysis

The mutation shifted the ratio of the HFA regioisomers (10-OH/13-OH) from 99:1 to 12:88

(Eser et al. 2020)

Lipoxygenases (LOX)

For the enzymatic production of 13R-hydroxy-docosahexaenoic acid and 13R,20-dihydroxy-docosahexaenoic acid from DHA

Catalytic mechanism-based site-directed mutagenesis

The catalytic properties of the mutant have shifted from 13S-LOX to 9R-LOX

(Yi et al. 2020)

Stability

Rhizopus oryzae lipase (ROL)

For the enzymatic production of TAGs

Sequence alignment

The mutant retains most of its activity at 70 °C, whereas the WT is incapable of functioning at temperatures above 60 °C

(Chow and Nguyen 2022)

Yarrowia lipolytica lipase Lip2

For the enzymatic production of MLM-SLs

Molecular dynamic (MD) simulation and the introduction of disulfide bonds

The mutant 4sN exhibited an increase in stability, with a rise in melting temperature (T_m) of 19.22 °C

(Li et al. 2022b)

Phospholipase C (PLC)

For enzymatic degumming of vegetable oils

B-factor analysis and MD simulation

The mutation F96R/Q153P showed the highest optimal reaction temperature (90 °C)

(Zhang et al. 2022b)

PLD

For the enzymatic modification of phospholipids

Disulfide bond engineering

The mutation showed a 3.1-fold increase in half-life (t_1/2) at 35 °C and a 5.7 °C rise in T_m

(Li et al. 2022a)