Efficient Hydrolysis of Gluten-Derived Celiac Disease-Triggering Immunogenic Peptides by a Bacterial Serine Protease from Burkholderia gladioli

Celiac disease is an autoimmune disorder triggered by toxic peptides derived from incompletely digested glutens in the stomach. Peptidases that can digest the toxic peptides may formulate an oral enzyme therapy to improve the patients’ health condition. Bga1903 is a serine endopeptidase secreted by Burkholderia gladioli. The preproprotein of Bga1903 consists of an N-terminal signal peptide, a propeptide region, and an enzymatic domain that belongs to the S8 subfamily. Bga1903 could be secreted into the culture medium when it was expressed in E. coli. The purified Bga1903 is capable of hydrolyzing the gluten-derived toxic peptides, such as the 33- and 26-mer peptides, with the preference for the peptide bonds at the carbonyl site of glutamine (P1 position). The kinetic assay of Bga1903 toward the chromogenic substrate Z-HPQ-pNA at 37 °C, pH 7.0, suggests that the values of Km and kcat are 0.44 ± 0.1 mM and 17.8 ± 0.4 s−1, respectively. The addition of Bga1903 in the wort during the fermentation step of beer could help in making gluten-free beer. In summary, Bga1903 is usable to reduce the gluten content in processed foods and represents a good candidate for protein engineering/modification aimed to efficiently digest the gluten at the gastric condition.

Oral Treatment With an Engineered Uricase, ALLN-346, Reduces Hyperuricemia, and Uricosuria in Urate Oxidase-Deficient Mice

Limitations in efficacy and/or tolerance of currently available urate-lowering therapies (ULTs), such as oral xanthine oxidase inhibitors, uricosurics, and intravenous uricase agents contribute to the development of refractory gout. Renal excretion is the major route of uric acid elimination, but the intestinal tract plays an increasingly recognized role in urate homeostasis, particularly in chronic kidney disease (CKD) in which the renal elimination of urate is impaired. We targeted intestinal degradation of urate in vivo with ALLN-346, an orally administered, engineered urate oxidase, optimized for proteolytic stability, and activity in the gut. We tested ALLN-346 in uricase/urate oxidase deficient mice (URKO mice) with severe hyperuricemia, hyperuricosuria, and uric acid crystalline obstructive nephropathy. A total of 55 male and female URKO mice were used in the two consecutive studies. These seminal, proof-of-concept studies aimed to explore both short- (7-day) and long-term (19-day) effects of ALLN-346 on the reduction of plasma and urine urate. In both the 7- and 19-day studies, ALLN-346 oral therapy resulted in the normalization of urine uric acid excretion and a significant reduction of hyperuricemia by 44 and 28% when therapy was given with food over 24 h or was limited for up to 6 h, respectively. Fractional excretion of uric acid (FEUA) was normalized with ALLN-346 therapy. Oral enzyme therapy with engineered urate oxidase (ALLN-346) designed to degrade urate in the intestinal tract has the potential to reduce hyperuricemia and the renal burden of filtered urate in patients with hyperuricemia and gout with and without CKD.

Effects of Exercise and Enzyme Therapy in Early Occupational Carpal Tunnel Syndrome: A Preliminary Study

Objectives. Occupational carpal tunnel syndrome (CTS) due to upper extremity overuse has in recent years been the most commonly recognized occupational disease in the Czech Republic and its prevalence has steadily increased. This pilot observation aimed to assess the effects of exercise techniques and oral enzyme therapy in automotive plant workers with early CTS. Patients and Methods. The observation comprised automotive plant assembly line workers in whom nerve conduction study revealed incipient CTS. The subjects were divided into three groups: a group practicing exercise techniques (exercising; N=15), a group receiving oral enzyme therapy (N=16), and a group of controls (N=14). Subjects in the control group were only observed without any specific intervention, which is a common procedure in incipient CTS. Throughout 9-week observation, the workers did their jobs. Prior to and after that period, the workers’ CTS-related symptoms were ascertained through structured interviews with a physician and the following median nerve parameters were measured: sensory conduction velocity (SCV) and distal motor latency (DML). Results. In both the exercise and enzyme therapy groups, statistically significant decreases in the total score for symptoms were achieved (p<0.0001), unlike controls. On final examination, both treated groups demonstrated significantly increased SCV as compared with the initial values (p=0.00013 and p<0.0001, respectively); in controls, the mean SCV did not significantly change. Similarly, a statistically significant shortening of DML was noted in the enzyme therapy group (p=0.008). Conclusion. The results showed the efficiency of both exercise and oral enzyme therapy in incipient CTS. These methods may be recommended for preventing more severe forms of CTS.