Numerous examples have been presented of enzyme activities, assayed in vitro, that appear relevant to the synthesis of structural polysaccharides, and to their assembly and subsequent degradation in the primary cell walls (PCWs) of higher plants. The accumulation of the corresponding mRNAs, and of the (immunologically recognized) proteins, has often also (or instead) been reported. However, the presence of these mRNAs, antigens and enzymic activities has rarely been shown to correspond to enzyme action in the living plant cell. In some cases, apparent enzymic action is observed in vivo for which no enzyme activity can be detected in in-vitro assays; the converse also occurs. Methods are reviewed by which reactions involving structural wall polysaccharides can be tracked in vivo. Special attention is given to xyloglucan endotransglucosylase (XET), one of the two enzymic activities exhibited in vitro by xyloglucan endotransglucosylase/hydrolase (XTH) proteins, because of its probable importance in the construction and restructuring of the PCW's major hemicellulose. Attention is also given to the possibility that some reactions observed in the PCW in vivo are not directly enzymic, possibly involving the action of hydroxyl radicals. It is concluded that some proposed wall enzymes, for example XTHs, do act in vivo, but that for other enzymes this is not proven. Contents I. Primary cell walls: composition, deposition and roles 642 II. Reactions that have been proposed to occur in primary cell walls 645 III. Tracking the careers of wall components in vivo: evidence for action of enzymes in the walls of living plant cells 656 IV. Evidence for the occurrence of nonenzymic polymer scission in vivo? 666 VI. Conclusion 667 References 667.

https://nph.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1469-8137.2004.00980.x

Primary cell wall metabolism: tracking the careers of wall polymers in living plant cells

β Galactosidases have been obtained from microorganisms such as fungi, bacteria and yeasts; plants, animals cells, and from recombinant sources. The enzyme has two main applications; the removal of lactose from milk products for lactose intolerant people and the production of galactosylated products. In order to increase their stability, reusability, and use in continuous reactors, these enzymes have been immobilized on both organic and inorganic support via adsorption, covalent attachment, chemical aggregation, microencapsulation, and entrapment. Free and immobilized preparations of β galactosidases have been exploited in various applications such as industrial, biotechnological, medical, analytical, and in different other applications. β galactosidase is widely used in food industry to improve sweetness, solubility, flavor, and digestibility of dairy products. Immobilized β galactosidases are employed for the continuous hydrolysis of lactose from whey and milk in a number of reactors such as hollow fiber reactors, tapered column reactors, packed bed reactors, fluidized bed reactors etc.

β Galactosidases and their potential applications: a review

Plant glycoside hydrolases involved in cell wall polysaccharide degradation.

The mechanisms controlling seed dormancy in Arabidopsis (Arabidopsis thaliana) have been characterized by proteomics using the dormant (D) accession Cvi originating from the Cape Verde Islands. Comparative studies carried out with freshly harvested dormant and after-ripened non-dormant (ND) seeds revealed a specific differential accumulation of 32 proteins. The data suggested that proteins associated with metabolic functions potentially involved in germination can accumulate during after-ripening in the dry state leading to dormancy release. Exogenous application of abscisic acid (ABA) to ND seeds strongly impeded their germination, which physiologically mimicked the behavior of D imbibed seeds. This application resulted in an alteration of the accumulation pattern of 71 proteins. There was a strong down-accumulation of a major part (90%) of these proteins, which were involved mainly in energetic and protein metabolisms. This feature suggested that exogenous ABA triggers proteolytic mechanisms in imbibed seeds. An analysis of de novo protein synthesis by two-dimensional gel electrophoresis in the presence of [35S]-methionine disclosed that exogenous ABA does not impede protein biosynthesis during imbibition. Furthermore, imbibed D seeds proved competent for de novo protein synthesis, demonstrating that impediment of protein translation was not the cause of the observed block of seed germination. However, the two-dimensional protein profiles were markedly different from those obtained with the ND seeds imbibed in ABA. Altogether, the data showed that the mechanisms blocking germination of the ND seeds by ABA application are different from those preventing germination of the D seeds imbibed in basal medium.

Proteomic Analysis of Seed Dormancy in Arabidopsis

The sensitive to freezing2-1 (sfr2-1) mutation causes freezing sensitivity in Arabidopsis thaliana. By mapping, transgenic complementation, and sequencing, sfr2-1 was revealed to be a mutation in gene At3g06510. A new knockout allele was obtained, and its identical freezing-sensitive phenotype confirmed that the SFR2 gene product is essential for freezing tolerance. Transcription of SFR2 was observed to be constitutive rather than stress inducible and was distributed throughout most aerial tissues. SFR2 encodes a protein homologous to family 1 glycosyl hydrolases (β-glycosidases), but the predicted AtSFR2 protein is divergent from all other family 1 β-glycosidases of Arabidopsis, showing closer homology to the sequences of several β-glycosidases from thermophilic archea and bacteria. After purification from a heterologous expression system, AtSFR2 displayed a specific hydrolytic activity against β-d-glucosides.

Kuan Chung Chen

Papers

Purification and characterization of isoforms of β-galactosidases in mung bean seedlings