The Preparation Of Coconut Fiber Immobilized Alpha-amylase And The Starch Dynamics Research Of High Temperature Hydrolysis

In this article, the coconut crude fiber is selected as the raw materal. The hemicellulose on the surface of coconut crube fiber is removed by heating dilute sulphuric acid hydrolysis, the coconut fiber riched in aldehyde was selective oxidation by sodium periodate, was made into immobilized carrier materials. Immobilized carrier material and liquid resistance to high temperature of alpha amylase reactions generated garment of coconut fiber immobilized by covalent bonds in combination with high temperature resistant alpha-amylase. By the immobilized enzyme hydrolyzing starch, we study the connection among coconut fibre immobilized high-temperature alpha enzyme, substrate concentration and reaction rate. liquid of coconut fiber immobilized high-temperature,and the relationship between the enzyme stability and dynamics of the process.Fresh coconut fiber surface is adhered with some loose structure of coconut chaff, the main components of coconut bran are hemicellulose and pentosan, half of these fibers is not stable, removed by heating by dilute sulfuric acid hydrolysis. The optimum process conditions: the sulfuric acid concentration is 1.3 mol/L, hydrolysis temperature is 102?,the hydrolysis time 2 h. By detecting the content of total reducing sugar in the hydrolysate,we can examine the extent of the coconut fiber hemicellulose on the surface removed.Coconut fiber surface treated by acid hydrolysis and washed is smooth and maroon, and it can be used to be as immobilized carrier materials of high temperature resistance of alpha amylase. In detail, selected the glucose as standard, DNS as chromogenic agent, using,coconut dress half a crude fiber surface fiber removal rate was measured 17.2 % by spectrophotometry.Sodium periodate was used as oxidant to oxidize the hydroxyl groups on the coir fiber selectively, and to make the coir fiber into a coir aldehyde fiber. During process of coir fiber oxidation, the hydroxylamine hydrochloride titrationwas taken to determine the concentration of aldehyde, in order to investigate the coir fiber oxidation process. The results showed that: when the concentration of sodium periodate solution is 0.7 mol/L,oxidation temperature is 50?, oxidation time is 3.5 h, and the pH of sodium periodate solution is 2.8, the oxidation effect is best and the amount of aldehyde group in the coir fiber increases, what's more, the structure and function of coir fiber will change, which will proved an effective response for next step synthetic new coir fiber polymer.The high temperature resistant alpha amylase can be obtained by Schiff reaction between the appropriate dilution of high temperature resistant alpha amylase and coconut aldehyde fiber, the optimum technological conditions: immobilized temperature is 55?,immobilization time is 18 h, buffer solution pH is 6.5, the original liquid enzyme activity is 225 U/mL.The coconut fiber immobilized high-temperature alpha amylase activity was 22.21 U. at the same time,we also study the immobilized high-temperature storage of alpha amylase and the number of repeated use, the results showed that the immobilized enzyme stored in the refrigerator, for 5 days repetively hydrolyzed starch 5 times, enzyme activity is > 60 %.Even if it is stored for 10 days, repetively hydrolyzed starch 10 times, the activity of the enzyme is > 19.2 %.To investigate coconut fiber immobilized high-temperature alpha amylase hydrolysis kinetics characteristics of starch, this article select add quantity of enzyme, enzyme solution temperature, the concentration of soluble starch solution and buffer solution pH and enzymo lysis time as parameters, studying the effect of the hydrolysis of starch solution.The results showed that the soluble starch concentration is 4 mg/mL, hydrolysis temperature is 80?, buffer solution pH is 5.5, hydrolysis time is 50 mins, high.temperature resistant alpha amylase garment of coconut fiber immobilized is 0.1250 g. In this condition, hydrolysis effect is best, the determination of starch in hydrolysate of saccharification rate was 23.2 %. Coconut fibre immobilized high-temperature alpha amylase hydrolysis of starch, the substrate concentration and reaction rate follows the mie equation, namely the michaelis constant Km=22.836 mg/mL, Vm=0.262 mg/(mL · min),and the mie equation for v =(0.262s)/(s+22.836).