Ultrasound mediated enzymatic hydrolysis of cellulose and carboxymethyl cellulose
Ультразвуковая модуляция ферментативного гидролиза целлюлозы и карбоксиметилцеллюлозы
2013-08-07
SCID: 54.1/kv42wy5f
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Michaelis-Menten kineticscarboxymethyl cellulose (CMC)cellulose particle size effectsrecombinant Trichoderma reesei cellulaseultrasound-mediated hydrolysis
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Abstract (AI)
A recombinant Trichoderma reesei cellulase was used for the ultrasound-mediated hydrolysis of soluble carboxymethyl cellulose (CMC) and insoluble cellulose of various particle sizes. The hydrolysis was carried out at low intensity sonication (2.4-11.8 W cm(-2) sonication power at the tip of the sonotrode) using 10, 20, and 40% duty cycles. [A duty cycle of 10%, for example, was obtained by sonicating for 1 s followed by a rest period (no sonication) of 9 s.] The reaction pH and temperature were always 4.8 and 50°C, respectively. In all cases, sonication enhanced the rate of hydrolysis relative to nonsonicated controls. The hydrolysis of CMC was characterized by Michaelis-Menten kinetics. The Michaelis-Menten parameter of the maximum reaction rate Vmax was enhanced by sonication relative to controls, but the value of the saturation constant Km was reduced. The optimal sonication conditions were found to be a 10% duty cycle and a power intensity of 11.8 W cm(-2) . Under these conditions, the maximum rate of hydrolysis of soluble CMC was nearly double relative to control. In the hydrolysis of cellulose, an increasing particle size reduced the rate of hydrolysis. At any fixed particle size, sonication at a 10% duty cycle and 11.8 W cm(-2) power intensity improved the rate of hydrolysis relative to control. Under the above mentioned optimal sonication conditions, the enzyme lost about 20% of its initial activity in 20 min. Sonication was useful in accelerating the enzyme catalyzed saccharification of cellulose.
Key Findings
1
CMC hydrolysis followed Michaelis–Menten kinetics; sonication increased Vmax and decreased Km compared to controls.
2
For cellulose, larger particle size reduced hydrolysis rate, but sonication at 10% duty cycle and 11.8 W cm^-2 improved rates at each particle size.
3
Low-intensity sonication (2.4–11.8 W cm^-2) enhanced enzymatic hydrolysis rates of both soluble CMC and insoluble cellulose versus nonsonicated controls.
4
Optimal sonication conditions were a 10% duty cycle and 11.8 W cm^-2 power intensity, nearly doubling the maximum hydrolysis rate of soluble CMC.
5
Under optimal sonication, the enzyme retained most activity but lost about 20% of initial activity after 20 minutes.
Research Object
Cellulose (insoluble cellulose of various particle sizes) and soluble carboxymethyl cellulose (CMC) undergoing enzymatic hydrolysis with recombinant Trichoderma reesei cellulase under ultrasound
Research Subject
Effect of low-intensity ultrasound (varying power and duty cycle) on the enzymatic hydrolysis kinetics and rate (including Michaelis–Menten parameters Vmax and Km), enzyme activity loss, and particle-size dependence of saccharification
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2013-08-07
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