Analysis and application of a suite of recombinant endo-β(1,3)-d-glucanases for studying fungal cell walls

Table 3 Percentages of cell wall degradation with PfLam16A at different conditions

Buffer	pH	Temp. (^oC)	Enzyme weight (µg)	Incubation time (h)	% of cell wall degradation ^a
Buffer	pH	Temp. (^oC)	Enzyme weight (µg)	Incubation time (h)	Mean ± SD
Citrate/phosphate 100 mM	5.5	70	20	20	49.76 ± 0.18
Citrate/phosphate 100 mM	5.5	70	20	25	51.86 ± 0.48^b
MES 100 mM	5.5	70	20	20	54.79 ± 1.48^b
	5.5	70	20	25	54.40 ± 1.48^b
	5.5	70	40	20	50.64 ± 1.06^b
	5.5	70	40	25	53.03 ± 0.36^b
	6.5	70	20	36	45.00 ± 0.11
Sodium acetate 100 mM	5	70	5	20	47.58 ± 0.05
	5	70	5	25	51.55 ± 0.32^b
	5	70	10	20	55.54 ± 0.21^b
	5	70	10	25	57.87 ± 0.49
	5	70	20	5	44.13 ± 1.44
				10	52.50 ± 1.58^b
				15	56.58 ± 0.53^b
				20	58.37 ± 1.82
				25	59.19 ± 2.82
				36	67.58 ± 0.21
	5	70	40	20	57.73 ± 3.30
	5	70	40	25	62.21 ± 4.01
Sodium phosphate 50 mM	6	60	20	15	33.25 ± 2.48
				20	33.39 ± 0.67
				25	39.17 ± 0.18
	6	70	20	15	39.67 ± 0.94
				20	44.84 ± 0.73
				25	48.90 ± 0.29
	6.5	60	20	15	30.53 ± 0.64
				20	31.95 ± 1.01
				25	37.53 ± 0.36
	6.5	70	20	15	38.89 ± 0.38
				20	42.68 ± 0.79
				25	45.38 ± 0.21
	7	70	20	15	41.59 ± 3.06
				20	39.82 ± 1.62
				25	41.14 ± 0.42
Sodium phosphate 100 mM	6	60	20	15 20 25	40.03 ± 0.98
					40.91 ± 1.28
					43.01 ± 0.22
	6	70	20	15 20 25	45.57 ± 0.97
					50.62 ± 3.59^b
					52.34 ± 3.76^b
	6	70	40	20 25	51.57 ± 6.52^b
	6	70	40	20 25	55.17 ± 5.78^b
	6.5	60	20	15	32.94 ± 2.08
				20	36.98 ± 2.05
				25	35.11 ± 0.86
	6.5	70	20	15	47.39 ± 3.99
				20	52.80 ± 3.32^b,c
				25	51.27 ± 2.79^b
	6.5	70	40	15	51.01 ± 4.19^b
				20	50.77 ± 0.28^b
				25	51.32 ± 0.69^b
				36	56.37 ± 2.33^b

^aPercentage of cell wall degradation calculated from at least two independent experiments
^bIn bold are shown the values and conditions of percentage of cell wall degradation similar to that of Quantazyme (50–56%)
^cUnderlined is shown the selected condition for the standard protocol of cell wall degradation (lowest enzyme amount and incubation time using the reaction conditions specified by the supplier)

ISSN: 1475-2859