Optimization of Lipase production from a novel strain Thalassospira permensis M35-15 using Response Surface Methodology

Figures & data

Table 1. Plackett–Burman design matrix and lipase activity of Thalassospira permensis M35-15.

Run	X1	X2	X3	X4	X5	X6	X7	X8	X9	X10	X11	Lipase activity(U)
1	1	−1	1	1	−1	1	−1	−1	−1	1	1	1.52
2	1	−1	1	−1	−1	−1	1	1	1	−1	1	1.23
3	−1	1	1	−1	1	−1	−1	−1	1	1	1	3.36
4	1	1	−1	1	−1	−1	−1	1	1	1	−1	4.03
5	−1	1	1	1	−1	1	1	−1	1	−1	−1	1.9
6	1	1	−1	1	1	−1	1	−1	−1	−1	1	48.26
7	−1	−1	−1	1	1	1	−1	1	1	−1	1	20.03
8	1	1	1	−1	1	1	−1	1	−1	−1	−1	20.39
9	−1	−1	−1	−1	−1	−1	−1	−1	−1	−1	−1	1.05
10	−1	1	−1	−1	−1	1	1	1	−1	1	1	2.34
11	1	−1	−1	−1	1	1	1	−1	1	1	−1	3.18
12	−1	−1	1	1	1	−1	1	1	−1	1	−1	4.01

Table 2. Results of significant influence by analysis of t test using Plackett–Burman design.

Variable	Effect	S.E	t	P
Intercept	9.275	3.133	2.96	0.060
X1	3.827	3.133	1.22	0.309
X2	4.105	3.133	1.31	0.281
X4	4.017	3.133	1.28	0.290
X5	7.263	3.133	2.32	0.103
X7	0.878	3.133	0.28	0.797
X8	−0.603	3.133	−0.19	0.860
X10	−6.202	3.133	−1.98	0.142
X11	3.515	3.133	1.12	0.344

Table 3. The design matrix of CCD with 4 various ingredients at different concentration.

	A: Glucose	B: Peptone	C: Yeast powder	D:Olive oil emulsifier	Lipase activity (U)
Run	(g/l)	(g/l)	(g/l)	(g/l)	Observed	Predicted
1	−1(4)	−1(8)	−1(3)	−1(20)	9.01 ± 0 .09	9.063
2	1(8)	−1(8)	−1(3)	−1(20)	8.17 ± 0 .14	8.059
3	−1(4)	1(12)	−1(3)	−1(20)	10.78 ± 0 .23	10.861
4	1(8)	1(12)	−1(3)	−1(20)	9.84 ± 0 .17	10
5	−1(4)	−1(8)	−1(3)	1(40)	9.62 ± 0 .04	9.414
6	1(8)	−1(8)	−1(3)	1(40)	8.45 ± 0 .06	8.658
7	−1(4)	1(12)	−1(3)	1(40)	11.05 ± 0 .07	11.37
8	1(8)	1(12)	−1(3)	1(40)	10.89 ± 0 .14	10.756
9	−1(4)	−1(8)	1(7)	−1(20)	8.41 ± 0 .07	8.569
10	1(8)	−1(8)	1(7)	−1(20)	8.19 ± 0 .25	8.138
11	−1(4)	1(12)	1(7)	−1(20)	9.95 ± 0 .08	10.01
12	1(8)	1(12)	1(7)	−1(20)	9.49 ± 0 .15	9.721
13	−1(4)	−1(8)	1(7)	1(40)	9.06 ± 0 .16	9.168
14	1(8)	−1(8)	1(7)	1(40)	9.04 ± 0 .07	8.984
15	−1(4)	1(12)	1(7)	1(40)	10.63 ± 0 .06	10.766
16	1(8)	1(12)	1(7)	1(40)	10.51 ± 0 .15	10.725
17	−2(2)	0(10)	0(5)	0(30)	10.28 ± 0 .11	10.071
18	2(10)	0(10)	0(5)	0(30)	9.11 ± 0 .07	9.026
19	0(6)	−2(6)	0(5)	0(30)	6.74 ± 0 .02	6.835
20	0(6)	2(14)	0(5)	0(30)	10.76 ± 0 .09	10.373
21	0(6)	0(10)	0(5)	−2(10)	9.62 ± 0 .07	9.476
22	0(6)	0(10)	0(5)	2(50)	10.98 ± 0 .13	10.831
23	0(6)	0(10)	−2(1)	0(30)	10.56 ± 0 .17	10.521
24	0(6)	0(10)	2(9)	0(30)	10.25 ± 0 .14	9.996
25	0(6)	0(10)	0(5)	0(30)	10.91 ± 0 .05	10.863
26	0(6)	0(10)	0(5)	0(30)	10.97 ± 0 .08	10.863
27	0(6)	0(10)	0(5)	0(30)	10.92 ± 0 .14	10.863
28	0(6)	0(10)	0(5)	0(30)	10.85 ± 0 .08	10.863
29	0(6)	0(10)	0(5)	0(30)	10.87 ± 0 .09	10.863
30	0(6)	0(10)	0(5)	0(30)	10.57 ± 0 .07	10.863
31	0(6)	0(10)	0(5)	0(30)	10.95 ± 0 .22	10.863

Table 4. ANOVA results of the CCD design for T. permensis M35-15 lipase production calculated by Minitab16.0.

Variable	DF	SS	MS	F	P
Model	14	35.7537	2.5538	48.33	0
A	1	1.638	1.638	31	0
B	1	18.7797	18.7797	355.36	0
C	1	2.754	2.754	52.11	0
D	1	0.4134	0.4134	7.82	0.013
A*A	1	3.0863	3.0863	58.4	0
B*B	1	9.1213	9.1213	172.6	0
C*C	1	0.8987	0.8987	17.01	0.001
D*D	1	0.6522	0.6522	12.34	0.003
A*B	1	0.0203	0.0203	0.38	0.544
A*C	1	0.0613	0.0613	1.16	0.298
A*D	1	0.3278	0.3278	6.2	0.024
B*C	1	0.0248	0.0248	0.47	0.503
B*D	1	0.1278	0.1278	2.42	0.139
C*D	1	0.0613	0.0613	1.16	0.298
Residual	16	0.8456	0.0528
Lack of fit	10	0.7350	0.0735	3.99	0.052
Pure error	6	0.1105	0.0184
Total	30	36.5992

Table 5. Significance of the CCD design for T. permensis M35-15 lipase production.

Variable	Coefficient estimate	Standard error	t	P
Intercept	10.8629	0.08689	125.021	0
A	−0.2613	0.04693	−5.567	0
B	0.8846	0.04693	18.851	0
C	0.3387	0.04693	7.219	0
D	−0.1312	0.04693	−2.797	0.013
A*A	−0.3285	0.04299	−7.642	0
B*B	−0.5648	0.04299	−13.138	0
C*C	−0.1773	0.04299	−4.124	0.001
D*D	−0.151	0.04299	−3.513	0.003
A*B	0.0356	0.05747	0.62	0.544
A*C	0.0619	0.05747	1.077	0.298
A*D	0.1431	0.05747	2.49	0.024
B*C	0.0394	0.05747	0.685	0.503
B*D	−0.0894	0.05747	−1.555	0.139
C*D	0.0619	0.05747	1.077	0.298

Figure 1. Response surface of lipase production effected by glucose and yeast powder.

Figure 2. Response surface of lipase production effected by peptone and olive oil emulsifier.

Table 6. Medium composition at different concentration for lipase secretion using Plackett-Burman experimental design.

Code	Variable	Unit	Levels +1	Levels −1
X1	Fructose	g l^−1	3	1
X2	Glucose	g l^−1	3	1
X3	Dummy	—	—	—
X4	Peanut oil emulsifier	g l^−1	40	20
X5	olive oil emulsifier	g l^−1	40	20
X6	Dummy	—	—	—
X7	Ammonium chloride	g l^−1	8	4
X8	Peptone	g l^−1	1	5
X9	Dummy	—	—	—
X10	Yeast powder	g l^−1	5	2
X11	Ammonium sulfate	g l^−1	8	4