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Table 1 Kinetic parameters of WT and H208F mutant

From: Engineering broad-spectrum digestion of polyuronides from an exolytic polysaccharide lyase

 

WT

H208F

k CAT (s−1)

K M (mM)

k CAT/K M

k CAT (s−1)

K M (mM)

k CAT/K M

Alginate

34.8

0.67

52.2

14.6

1.35

10.8

PolyManA

62.2

0.41

151.8

21.3

0.41

51.5

Poly-GulA

34.6

5.99

5.8

1.4

2.22

0.7

Poly-MG

22.2

0.57

38.7

2.0

0.98

2.1

Poly-GlcA

0.4

1.40

0.3

6.1

0.57

10.6

  1. WT and H208F mutant were added to 10 different solutions containing 7.8–4000 µg/mL substrate in 20 mM sodium phosphate buffer at pH 8.5 for alginate-based polysaccharides, and pH 8 for poly-GlcA. Enzymatic activity was monitored by absorbance at 235 nm, and all reactions were performed in triplicate. Turnover rate (k CAT) was calculated using an extinction coefficient of 6150 M−1 cm−1 for the unsaturated product [11, 12]. K M values are based upon an effective molecular weight of 176 for the monosaccharide unit, the smallest product formed by Smlt2602 during catalysis [12, 52]
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Biotechnology for Biofuels and Bioproducts

ISSN: 2731-3654

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