By R.J. Sturgeon
In recent times the main major advances in carbohydrate examine were made within the wisdom of the constitution and serve as of carbohydrates within the macromolecular country. This identify addresses these parts of the topic during which the authors think crucial paintings is being conducted.
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Extra info for Advances in Macromolecular Carbohydrate Research, Volume 1
These substrates include carboxymethyl debranched arabinan, Procion Brilliant Red linear arabinan, and Remazol Brilliant Blue-or-Black-CM-linear arabinan. For practical applications we prefer the substrate dyed with Procion Red. After incubation with enzyme the unreacted substrate is precipitated in 75% ethanol and the dyed hydrolysis products are measured at 520 nm. Advanced methods to identify endo- or ejo-acting arabinanases have been introduced by the use of fast and high performance liquid chromatography (FPLC and HPLC.
Probably a carboxyl group and a histidine imidazolium group are involved with the enzyme substrate complex and influence the activity . There is a cooperative catalytic action of a proton donor, being the imidazolium ion, and a nucleophile in the form of a carboxyl group. Probably this is a two-step mechanism which includes the formation of a glycosyl—enzyme intermediate. The enzyme also catalyzes a ^ra«5-glycosylation Table 9. (M) Activity towards:^ />-Nitrophenyl-p-L-arabinopyranoside /7-Nitrophenyl-a-D-galactopyranoside p-Nitrophenyl-P-D-galactopyranoside p-Nitrophenyl-a-D-fucopyranoside Note: ^+ activity; - no activity.
Advanced methods to identify endo- or ejo-acting arabinanases have been introduced by the use of fast and high performance liquid chromatography (FPLC and HPLC. respectively). These methods have been used to develop a rapid screening procedure for arabinanases in crude mixtures of polysaccharidases, but are also applicable for xylanases . 2 c o GL - 30 min. 1i CLI Q 2- 15 min. A J - ^5 min. ^ c) A 1 JJi J 1 ll nn 1 UhiH.... s^ 1 10 10 20 30 40 5( retention time (min) Figure 5. Analysis of an enc/o-arabinanase digest of a linear arabinan by HPLC (Carbopac PA 1; sodium acetate gradient; pulsed amperometric detection).