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Literature summary for 3.2.1.B1 extracted from

  • Shi, C.; Lu, X.; Ma, C.; Ma, Y.; Fu, X.; Yu, W.
    Enhancing the thermostability of a novel beta-agarase AgaB through directed evolution (2008), Appl. Biochem. Biotechnol., 151, 51-59.
    View publication on PubMed
Search for more literature for 3.2.1.B1

Protein Variants

Protein Variants Comment Organism
L122Q mutant enzyme shows similar thermostability with wild-type AgaB. Specific activity 1.3fold higher than that of wild-type enzyme Pseudoalteromonas sp.
additional information to increase the thermostability of beta-agarase AgaB by directed evolution, the mutant gene libraries are generated by error-prone polymerase chain reaction and deoxyribonucleic acid shuffling. A mutant S2 is obtained through two rounds of error-prone polymerase chain reaction and a single round of DNA shuffling and selection. It has higher thermostability and slightly increased catalytic activity than wild-type AgaB. Melting temperature (Tm) of S2, as determined by circular dichroism, is 4.6°C higher than that of wild-type AgaB, and the half-life of S2 is 350 min at 40°C, which is 18.4-fold longer than that of the wild-type enzyme. Saturation mutagenesis and hydrophobic cluster analysis indicate that hydrophobic interaction might be the key factor that enhances the enzyme stability Pseudoalteromonas sp.
N446I mutant enzyme has enhanced thermostability. 10-20% decrease of the specific activity compared to wild-type enzyme Pseudoalteromonas sp.
N446L half-life of mutant enzyme at 40°C is 12.9fold longer than that of wild-type AgaB Pseudoalteromonas sp.
N446V half-life of mutant enzyme at 40°C is 18.2fold longer than that of wild-type AgaB Pseudoalteromonas sp.

Organism

Organism UniProt Comment Textmining
Pseudoalteromonas sp.
-
 expressed in Escherichia coli
-

Synonyms

Synonyms Comment Organism
AgaB
-
 Pseudoalteromonas sp.
beta-agarase AgaB
-
 Pseudoalteromonas sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
40
-
 optimal reaction temperature of mutant enzyme S2 and wild-type enzyme Pseudoalteromonas sp.

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
35
-
 wild-type AgaB has low thermostability when temperature is above 35°C Pseudoalteromonas sp.
40
-
 half-life of mutant S2 is 350 min, which was 18.4fold longer than that of wild-type AgaB Pseudoalteromonas sp.
49
-
 melting temperature of wild-type enzyme is 49.2°C Pseudoalteromonas sp.
54
-
 melting temperature of mutant enzyme S2 is 53.8°C Pseudoalteromonas sp.