Biotransformação otimizada de hidrolisado de casca de melancia tratada com ácido em etanol

Chaudhary, A.; Akram, A. M.; Ahmad, Qurat-ul-Ain; Hussain, Z.; Zahra, S.; Minahal, Q.; Azhar, S.; Ahmad, S.; Hayat, S.; Javed, M. A.; Haider, M. S.; Ali, Q.; Karita, S.

A. Chaudhary ^**e-mail: asma.ch@ue.edu.pk; saim1692@gmail.com

University of Education, Department of Zoology, Division of Science and Technology, Lahore, Pakistan

http://orcid.org/0000-0001-6837-7611

A. M. Akram

University of Education, Department of Zoology, Division of Science and Technology, Lahore, Pakistan

http://orcid.org/0000-0003-2610-2083

Qurat-ul-Ain Ahmad

University of Education, Department of Zoology, Division of Science and Technology, Lahore, Pakistan

http://orcid.org/0000-0003-2976-1842

Z. Hussain

University of Education, Department of Zoology, Division of Science and Technology, Lahore, Pakistan

http://orcid.org/0000-0002-0965-2051

S. Zahra

University of Education, Department of Physics, Division of Science and Technology, Lahore, Pakistan

http://orcid.org/0000-0001-8901-6639

Q. Minahal

University of Education, Department of Zoology, Division of Science and Technology, Lahore, Pakistan

S. Azhar

University of the Punjab, Institute of Zoology, Lahore, Pakistan

S. Ahmad

University of the Punjab Lahore, Department of Entomology, Lahore, Pakistan

S. Hayat

The University of Lahore, Institute of Molecular Biology and Biotechnology, Lahore, Pakistan

M. A. Javed

University of the Punjab Lahore, Department of Plant Breeding and Genetics, Lahore, Pakistan

http://orcid.org/0000-0001-9629-4215

M. S. Haider

University of the Punjab Lahore, Department of Plant Pathology, Lahore, Pakistan

Q. Ali ^**e-mail: asma.ch@ue.edu.pk; saim1692@gmail.com

The University of Lahore, Institute of Molecular Biology and Biotechnology, Lahore, Pakistan

http://orcid.org/0000-0002-3160-4830

S. Karita

Mie University, Graduate School of Bioresources, Tsu city, Mie, Japan

http://orcid.org/0000-0003-2733-816X

*e-mail: asma.ch@ue.edu.pk; saim1692@gmail.com

Fermentation Parameters	Coded variables
Fermentation Parameters	Coded variable	Minimum variable	Middle variable	Maximum variable
WMPH (mL)	X₁	25	50	75
Incubation duration (days)	X₂	1	8	15
Incubation Temperature (°C)	X₃	25	32.50	40

Experimental Runs	WMPH	Incubation days	Temperature °C
1	50	8	19.89
2	50	8	45.11
3	50	19.77	32.5
4	50	3.77	32.5
5	50	8	32.5
6	92.04	8	32.5
7	75	1	25
8	25	15	25
9	50	8	32.5
10	25	1	25
11	50	8	32.5
12	50	8	32.5
13	7.96	8	32.5
14	25	1	40
15	50	8	32.5
16	25	15	40
17	75	1	40
18	75	15	40
19	75	15	25
20	50	8	32.5

WMP composition	Contents
Total sugars	92.9±0.03
Reducing sugars	25.8±0.06
Total lipids	3.4±0.01
Total proteins	3.9±0.02
Extractives*	20.96±0.05
Soluble lignin*	10.35±0.29
Hemicellulose*	30.90±2.26
Cellulose+insoluble lignin^* * Crude cellulose + insoluble lignin (%) = 100 – (Extractive % + Hemicellulose % + Lignin %).	37.79±0.31
Moisture contents	8.3±0.04
Weight loss	17.99±0.47

	Parameters			S. cerevisiae K7			M. cibodasensis Y34
Experimental Runs	WMP	Incubation days	Temp °C	Ethanol Yield g/g	Ethanol Titer g/L	Growth (O.Ds)	Ethanol Yield g/g	Ethanol Titer g/L	Growth (O.Ds)
1	75	15	40	23.6±0.03	19.6±0.03	4.0±0.03	27.4±0.06	24.8±0.21	2.6±0.02
2	50	8	32.5	27.8±0.06	23.9±0.01	3.9±0.03	28.7±0.06	26.2±0.10	2.5±0.04
3	75	15	25	31.3±0.03	31.2±0.04	0.1±0.09	35.4±0.36	35.3±0.03	0.1±0.03
4	75	1	40	33.0±0.03	32.9±0.04	0.1±0.13	34.3±0.06	32.5±0.03	1.8±0.03
5	25	1	40	10.9±0.09	6.9±0.18	4.0±0.11	11.1±0.06	7.6±0.11	3.5±0.19
6	50	8	32.5	28.0±0.06	23.9±0.01	4.1±0.02	27.3±0.06	23.4±0.03	3.9±0.02
7	25	15	40	19.4±0.09	19.3±0.11	0.1±0.02	19.7±0.06	19.6±0.06	0.1±0.02
8	7.96	8	32.5	15.5±0.04	15.3±0.05	0.2±0.08	16.1±0.01	15.7±0.02	0.4±0.10
9	50	8	32.5	27.5±0.06	23.9±0.01	3.6±0.05	27.4±0.06	23.2±0.07	4.2±0.04
10	50	8	19.89	24.5±0.06	15.6±0.07	8.9±0.03	25.8±0.06	22.3±0.10	3.5±0.05
11	50	19.77	32.5	24.0±0.06	17.6±0.06	6.4±0.03	20.3±0.06	17.8±0.07	2.5±0.01
12	50	8	45.11	26.1±0.06	25.4±0.01	0.7±0.03	28.2±0.06	27.8±0.02	0.4±0.03
13	50	8	32.5	25.3±0.06	23.9±0.01	1.4±0.05	27.8±0.06	23.7±0.06	4.1±0.08
14	50	3.77	32.5	25.4±0.06	14.7±0.23	10.7±0.15	25.4±0.06	19.4±0.08	6.0±0.04
15	92.04	8	32.5	31.7±0.16	26.3±0.18	5.4±0.05	35.4±0.05	32.9±0.09	2.5±0.09
16	50	8	32.5	28.4±0.06	23.9±0.01	4.5±0.04	28.4±0.06	24.7±0.08	3.7±0.03
17	75	1	25	23.0±0.03	15.8±0.05	7.2±0.09	29.6±0.06	15.9±0.29	13.7±0.28
18	25	1	25	18.9±0.09	17.4±0.04	1.5±0.03	19.7±0.06	8.1±0.12	11.6±0.19
19	25	15	25	27.6±0.04	27.5±0.02	0.1±0.02	28.1±0.06	28.0±0.05	0.1±0.14
20	50	8	32.5	28.3±0.06	23.9±0.01	4.4±0.02	28.6±0.06	26.7±0.02	1.9±0.11

	Fermentation Conditions			S. cerevisiae K7			M. cibodasensis(Y34)
Experimental Runs	WMPH (mL)	Incubation days	Temp °C	Total RS (g/L)	Consumed RS (g/L)	Remaining RS (g/L)	Total RS (g/L)	Consumed RS (g/L)	Remaining RS (g/L)
1	75	15	40	23.6±0.03	19.6±0.03	4.0±0.03	27.4±0.06	24.8±0.21	2.6±0.02
2	50	8	32.5	27.8±0.06	23.9±0.01	3.9±0.03	28.7±0.06	26.2±0.10	2.5±0.04
3	75	15	25	31.3±0.03	31.2±0.04	0.1±0.09	35.4±0.36	35.3±0.03	0.1±0.03
4	75	1	40	33.0±0.03	32.9±0.04	0.1±0.13	34.3±0.06	32.5±0.03	1.8±0.03
5	25	1	40	10.9±0.09	6.9±0.18	4.0±0.11	11.1±0.06	7.6±0.11	3.5±0.19
6	50	8	32.5	28.0±0.06	23.9±0.01	4.1±0.02	27.3±0.06	23.4±0.03	3.9±0.02
7	25	15	40	19.4±0.09	19.3±0.11	0.1±0.02	19.7±0.06	19.6±0.06	0.1±0.02
8	7.96	8	32.5	15.5±0.04	15.3±0.05	0.2±0.08	16.1±0.01	15.7±0.02	0.4±0.10
9	50	8	32.5	27.5±0.06	23.9±0.01	3.6±0.05	27.4±0.06	23.2±0.07	4.2±0.04
10	50	8	19.89	24.5±0.06	15.6±0.07	8.9±0.03	25.8±0.06	22.3±0.10	3.5±0.05
11	50	19.77	32.5	24.0±0.06	17.6±0.06	6.4±0.03	20.3±0.06	17.8±0.07	2.5±0.01
12	50	8	45.11	26.1±0.06	25.4±0.01	0.7±0.03	28.2±0.06	27.8±0.02	0.4±0.03
13	50	8	32.5	25.3±0.06	23.9±0.01	1.4±0.05	27.8±0.06	23.7±0.06	4.1±0.08
14	50	3.77	32.5	25.4±0.06	14.7±0.23	10.7±0.15	25.4±0.06	19.4±0.08	6.0±0.04
15	92.04	8	32.5	31.7±0.16	26.3±0.18	5.4±0.05	35.4±0.05	32.9±0.09	2.5±0.09
16	50	8	32.5	28.4±0.06	23.9±0.01	4.5±0.04	28.4±0.06	24.7±0.08	3.7±0.03
17	75	1	25	23.0±0.03	15.8±0.05	7.2±0.09	29.6±0.06	15.9±0.29	13.7±0.28
18	25	1	25	18.9±0.09	17.4±0.04	1.5±0.03	19.7±0.06	8.1±0.12	11.6±0.19
19	25	15	25	27.6±0.04	27.5±0.02	0.1±0.02	28.1±0.06	28.0±0.05	0.1±0.14
20	50	8	32.5	28.3±0.06	23.9±0.01	4.4±0.02	28.6±0.06	26.7±0.02	1.9±0.11

Responses	Source	Sum of Squares	DF	Mean Square	F value	p value
*Ethanol Yield (S. cerevisiae* K7)**	Model	0.17	9	0.019	8.21	0.002 Significant
	Residual	0.021	9	0.002
	Lack of Fit	0.021	5	0.004	60.48	0.008 Significant
	Pure Error	0.0002	4	0.00006
	Core Total	0.19	19
*Ethanol Yield (M. cibodasensis* Y34)**	Model	0.17	9	0.019	10.97	0.0006 Significant
	Residual	0.016	9	0.001
	Lack of Fit	0.015	5	0.003	48.93	0.0010 Significant
	Pure Error	0.0001	4	0.00006
	Core Total	0.19	19
*Ethanol Titer (S. cerevisiae* K7)**	Model	1.07	9	0.017	3.40	0.0412 Significant
	Residual	0.36	9	0.025
	Lack of Fit	0.31	5	0.063	209.69	<0.0001 Significant
	Pure Error	0.0012	4	0.0003
	Core Total	1.42	19
*Ethanol Titer (M. cibodasensis* Y34)**	Model	1.40	9	0.016	3.40	0.0414 Significant
	Residual	0.41	9	0.046
	Lack of Fit	0.40	5	0.080	31.15	0.0027 Significant
	Pure Error	0.010	4	0.002
	Core Total	1.89	19
*Yeast Growth (S. cerevisiae* K7)**	Model	1.64	9	0.18	24.13	<0.0001 Significant
	Residual	0.068	9	0.007
	Lack of Fit	0.064	5	0.013	13.20	0.0135 Significant
	Pure Error	0.003	4	0.0009
	Core Total	1.76	19
*Yeast Growth (M. cibodasensis* Y34)**	Model	1.78	9	0.20	12.58	0.0004 Significant
	Residual	0.14	9	0.016
	Lack of Fit	0.14	5	0.028	28.46	0.0032 Significant
	Pure Error	0.003	4	0.0009
	Core Total	1.98	19

Responses	C.V %	R-Squared	Adj R-Squared	Pred R-Squared	Adeq Precision
*Ethanol yield (S. cerevisiae* K7)**	26.51	0.8916	0.7832	0.1073	10.210
*Ethanol yield (M. cibodasensis* Y34)**	17.80	0.9164	0.8329	0.1074	12.537

Responses	Predicted Value	Experimental Value	Residual	Error (%)
*Ethanol yield (S. cerevisiae* K7)**	0.34	0.36±0.02	0.02	5.88
*Ethanol yield (M. cibodasensis* Y34)**	0.39	0.40±0.01	0.01	2.56
*Ethanol titer (S. cerevisiae* K7)**	9.5	9.4±0.03	-0.1	-1.057
*Ethanol titer (M. cibodasensis* Y34)**	11.4	11.5±0.04	0.1	0.87
*Yeast growth (S. cerevisiae* K7)**	0.65	2.50±0.05	1.85	28.4
*Yeast growth (M. cibodasensis* Y34)**	0.84	2.59±0.01	1.75	20.8

Y = β_{0} + β_{1} X_{1} + β_{2} X_{2} + β_{3} X_{3} + β_{11} X_{1}^{2} + β_{22} X_{2}^{2} + β_{33} X_{3}^{2} + β_{12} X_{1} X_{2} + β_{13} X_{1} X_{3} + β_{23} X_{2} X_{3} + e

[1] *e-mail: asma.ch@ue.edu.pk; saim1692@gmail.com

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Biotransformação otimizada de hidrolisado de casca de melancia tratada com ácido em etanol

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