CjMan26A

Ex-Man0107

(EC.3.2.1.78)endo-β-Mannanase

CAZy Family: GH26

PROPERTIES

1.ELECTROPHORETIC PURITY

-Single band on SDS-gel electrophoresis (MW ~48 kDa)

Figure 1. Electrophoresis analysis of CjMan26A. M, molecular weight marker (PageRuler Prestained Protein Ladder, Thermo Scientific); lane 1, culture lysate before IPTG induction; lane 2, culture lysate after IPTG induction; lane 3, CjMan26A purified from Ni sepharose fastflow column.

2.SPECIFIC ACTIVITY

ThepNP-β-manactivitywas not detected.

3.RELATIVE RATES OF HYDROLYSIS OF SUBSTRATES

Table 1.Biochemical Properties of CjMan26A^a

^aThe units of the kinetic parameterk_cat/K_Mfor the mannooligosaccharides are min^-1μM^-1. The units of the kinetic parameters for the polysaccharide substrates are as follows:k_cat, min-1;K_M, mg mL^-1;k_cat/K_M, mL min^-1mg^-1.

^bNot determined.

Figure 2. HPAEC detection of mannosidase hydrolysis of different mannoses.

Figure 3.Release of Man2 and Glc-Man from glucomannan by GH5 and GH26 mannanases. Glucomannan (2 mg/mL) was incubated with the indicated enzyme in 50 mM sodium phosphate/12 mM citrate buffer, and at timed intervals, the quantity ofMan2 and Glc-Man was determined by HPLC: (left) amounts of Man2 and Glc-Man released and (right) ratios of the two disaccharides [1].

4.PHYSICOCHEMICAL PROPERTIES

pH Optima: 7.0

Temperature Optima: 35°C

5.STORAGE CONDITIONS

The enzyme should be stored at -20°C. For assay, this enzyme should be diluted in phosphate buffer (50 mM) pH 7.0. Swirl to mix the enzyme immediately prior to use.

6. REFERENCES

[1] Tailford L E, Ducros M A, Flint J E, et al. Understanding How Diverse β-Mannanases Recognize Heterogeneous Substrates[J]. Biochemistry, 2009, 48(29): 7009-7018.