Enzymes

Lactose malabsorption.
Grand RJ, Montgomery RK.
Curr Treat Options Gastroenterol. 2008 Feb;11(1):19-25.

 Lactose malabsorption is a syndrome producing constellation of symptoms, including abdominal pain, bloating, flatulence, diarrhea, and sometimes nausea and/or vomiting. Primary causes of lactose malabsorption due to loss of intestinal lactase activity include genetic/racial lactase nonpersistence, congenital lactase deficiency, and developmental  lactase deficiency. Secondary lactose malabsorption can be caused by any disorder that injures the small intestinal mucosa, such as viral gastroenteritis, celiac disease, allergic (eosinophilic) gastroenteritis, and radiation enteritis. The diagnosis depends on careful clinical evaluation and is customarily confirmed with a lactose breath hydrogen test. As the symptoms are nonspecific, many adults diagnosed with lactose malabsorption actually have irritable bowel syndrome. Treatment consists of a trial of eliminating lactose-containing dairy foods, with supplementation of alternative calcium and protein sources. Commercial enzyme products containing β-galactosidases can be prescribed to help patients digest dietary lactose. Long-term lactose restriction usually is not necessary and can lead to reduced bone mineral density. PMID: 21063860

  Lactose intolerance in infants, children, and adolescents.
Heyman MB; Committee on Nutrition.
Pediatrics. 2006 Sep;118(3):1279-86.

 The American Academy of Pediatrics Committee on Nutrition presents an updated review of lactose intolerance in infants, children, and adolescents. Differences between primary, secondary, congenital, and developmental lactase deficiency that may result in lactose intolerance are discussed. Children with suspected lactose intolerance can be assessed clinically by dietary lactose elimination or by tests including noninvasive hydrogen breath testing or invasive intestinal biopsy determination of lactase (and other disaccharidase) concentrations. Treatment consists of use of lactase-treated dairy products or oral lactase supplementation, limitation of lactose-containing foods, or dairy elimination. The American Academy of Pediatrics supports use of dairy foods as an important source of calcium for bone mineral health and of other nutrients that facilitate growth in children and adolescents. If dairy products are eliminated, other dietary sources of calcium or calcium supplements need to be provided. PMID: 16951027

  A food-grade enzyme preparation with modest gluten detoxification properties.
Ehren J, Morón B, Martin E, Bethune MT, Gray GM, Khosla C.
PLoS One. 2009 Jul 21;4(7):e6313.

 Celiac sprue is a life-long disease characterized by an intestinal inflammatory response to dietary gluten. A gluten-free diet is an effective treatment for most patients, but accidental ingestion of gluten is common, leading to incomplete recovery or relapse. Food-grade proteases capable of detoxifying moderate quantities of dietary gluten could mitigate this problem. We evaluated the gluten detoxification properties of two food-grade enzymes, aspergillopepsin (ASP) from Aspergillus niger and dipeptidyl peptidase IV (DPPIV) from Aspergillus oryzae. The ability of each enzyme to hydrolyze gluten was tested against synthetic gluten peptides, a recombinant  gluten protein, and simulated gastric digests of whole gluten and whole-wheat bread. Reaction products were analyzed by mass spectrometry, HPLC, ELISA with a monoclonal antibody that recognizes an immunodominant gluten epitope, and a T cell proliferation assay. ASP markedly enhanced gluten digestion relative to pepsin, and cleaved recombinant alpha2-gliadin at multiple sites in a non-specific manner. When used alone, neither ASP nor DPPIV efficiently cleaved synthetic immunotoxic glutenpeptides. This lack of specificity for gluten was especially evident in the presence of casein, a competing dietary protein. However, supplementation of ASP with DPPIV enabled detoxification of moderate amounts of gluten in the presence of excess casein and in whole-wheat bread. ASP was also effective at enhancing the gluten-detoxifying efficacy of cysteine endoprotease EP-B2 under simulated gastric conditions. Clinical studies are warranted to evaluate whether a fixed dose ratio combination of ASP and DPPIV can provide near-term relief for celiac patients suffering from inadvertent  gluten exposure. Due to its markedly greater hydrolyticactivity against gluten than endogenous pepsin, food-grade ASP may also augment the activity of therapeutically relevant doses of glutenases such as EP-B2 and certain prolyl endopeptidases.
PMID: 19621078

  Intestinal digestive resistance of immunodominant gliadin peptides.
Hausch F, Shan L, Santiago NA, Gray GM, Khosla C.
Am J Physiol Gastrointest Liver Physiol. 2002 Oct;283(4):G996-G1003.

 Two recently identified immunodominant epitopes from alpha-gliadin account for most of the stimulatory activity of dietary gluten on intestinal and peripheral T lymphocytes in patients with celiac sprue. The proteolytic kinetics of peptides containing these epitopes were analyzed in vitro using soluble proteases from bovine and porcine pancreas and brush-border membrane vesicles from adult rat intestine. We showed that these proline-glutamine-rich epitopes are exceptionally resistant to enzymatic processing. Moreover, as estimated from the residual peptide structure and confirmed by exogenous peptidase supplementation, dipeptidyl peptidase IV and dipeptidyl carboxypeptidase I were identified as the rate-limiting enzymes in the digestive breakdown of these peptides. A similar conclusion also emerged from analogous studies with brush-border membrane from a human intestinal biopsy. Supplementation of rat brush-border membrane with trace quantities of a bacterial prolyl endopeptidase led to the rapid destruction of the immunodominant epitopes in these peptides. These results suggest a possible enzyme therapy strategy for celiac sprue, for which the only current therapeutic option is strict exclusion of gluten-containing food. PMID: 12223360

  Genetic evidence for role of DPP IV in intestinal hydrolysis and assimilation of prolyl peptides.
Tiruppathi C, Miyamoto Y, Ganapathy V, Leibach FH.
Am J Physiol. 1993 Jul;265(1 Pt 1):G81-9.

 The functional role of dipeptidyl peptidase IV (DPP IV) in the intestinal hydrolysis and assimilation of prolyl peptides was investigated using Japan F344 rats, which genetically lack this enzyme. USA F344 rats possess normal activity of this enzyme and served as matched controls. Intestinal brush-border membranes from the control rats were able to hydrolyze several proline-containing peptides. The hydrolytic ability of the brush-border membranes from the Japan rats against these peptides was markedly low. The difference in the hydrolytic activities between the two groups of rats was solely due to the absence of DPP IV in the Japan rats. There was no difference in the growth rate between the two groups of rats fed a reference diet whose protein constituents were not rich in proline. When the protein source was changed to gliadin, a proline-rich protein, USA F344 rats maintained their body weight for a 4-wk period on this diet, whereas the Japan rats experienced a significant weight loss under similar conditions. In situ perfusion experiments in intact animals revealed that the ability of morphiceptin (a peptide primarily hydrolyzable by DPP IV), when administered into the intestinal lumen, to block the cholera toxin-induced water secretion was significantly greater in Japan F344 rats than in USA F344 rats, indicating the resistance of morphiceptin to hydrolytic breakdown in the intestinal lumen of the Japan rats. It is concluded that the intestinal DPP IV plays a significant role in the hydrolysis of prolyl peptides and assimilation of proline-rich proteins. PMID: 8101699

 Reduction of non-digestible oligosaccharides in soymilk: application of engineered lactic acid bacteria that produce alpha-galactosidase.
LeBlanc JG, Silvestroni A, Connes C, Juillard V, de Giori GS, Piard JC, Sesma F.
Genet Mol Res. 2004 Sep 30;3(3):432-40.

 Human consumption of soy-derived products has been limited by the presence of non-digestible oligosaccharides (NDO), such as the alpha-galactooligosaccharides raffinose and stachyose. Most mammals, including man, lack pancreatic alpha-galactosidase (alpha-Gal), which is necessary for the hydrolysis of these sugars. However, such NDO can be fermented by gas-producing microorganisms present in the cecum and large intestine, which in turn can induce flatulence and other gastrointestinal disorders in sensitive individuals. The use of microorganisms expressing alpha-Gal is a promising solution to the elimination of NDO before they reach the large intestine. In the present study, lactic acid bacteria engineered to degrade NDO have been constructed and are being used as a tool to evaluate this solution. The alpha-Gal structural genes from Lactobacillus plantarum ATCC8014 (previously characterized in our laboratory) and from guar have been cloned and expressed in Lactococcus lactis. The gene products were directed to different bacterial compartments to optimize their possible applications. The alpha-Gal-producing strains are being evaluated for their efficiency in degrading raffinose and stachyose: i) in soymilk fermentation when used as starters and ii) in situ in the upper gastrointestinal tract when administered to animals orally, as probiotic preparations. The expected outcomes and possible complications of this project are discussed. PMID: 15614733

Does Beano prevent gas? A double-blind crossover study of oral alpha-galactosidase to treat dietary oligosaccharide intolerance.
Ganiats TG, Norcross WA, Halverson AL, Burford PA, Palinkas LA.
J Fam Pract. 1994 Nov;39(5):441-5.

 Beano, an over-the-counter oral solution of alpha-galactosidase, is used to prevent flatus and other gastrointestinal symptoms resulting from a high-fiber diet. The efficacy of this product, however, has not yet been adequately evaluated. Nineteen subjects were randomized into two groups and fed test meals of meatless chili. At the first test meal, group 1 received eight drops of alpha-galactosidase solution and group 2 received eight drops of placebo. After the meal, subjects were asked to keep a careful record of gastrointestinal symptoms, including occurrences of intestinal gas passage, for the next 6 hours. One week later, an identical test meal was served to each study subject and the solutions were reversed. Again subjects recorded their symptoms for the next 6 hours. Data were analyzed by means of paired t tests. The number of flatulence events per hour was significantly less in the group treated with alpha-galactosidase than placebo over the 6-hour follow-up period (F = 2.87, P = .016). When the two groups were compared at each follow-up interval, this difference was statistically significant only for the 5th hour after ingesting the test meal (t = 2.19, P = .04). No differences between the two groups were found in the extent of bloating or pain following the meal. Oral alpha-galactosidase solution is efficacious, at least in some patients, for the prophylaxis of gastrointestinal intolerance of oligosaccharides. PMID: 7964541

  The effect of oral alpha-galactosidase on intestinal gas production and gas-related symptoms.
Di Stefano M, Miceli E, Gotti S, Missanelli A, Mazzocchi S, Corazza GR.
Dig Dis Sci. 2007 Jan;52(1):78-83

 Bloating, abdominal distention, and flatulence represent very frequent complaints in functional disorders but their pathophysiology and treatment are largely unknown. Patients frequently associate these symptoms with excessive intestinal gas and the reduction of gas production may represent an effective strategy. The aim was to evaluate the effect of alpha-galactosidase administration, in a randomized double-blind placebo-controlled protocol, on intestinal gas production and gas-related symptoms after a challenge test meal in healthy volunteers. Eight healthy volunteers ingested 300 or 1200 GalU of alpha-galactosidase or placebo during a test meal containing 420 g of cooked beans. Breath hydrogen excretion and occurrence of bloating, abdominal pain, discomfort, flatulence, and diarrhea were measured for 8 hr. The administration of 1200 GalU of alpha-galactosidase induced a significant reduction of both breath hydrogen excretion and severity of flatulence. A reduction in severity was apparent for all considered symptoms, but both 300 and 1200 GalU induced a significant reduction in the total symptom score. Alpha-galactosidase reduced gas production following a meal rich in fermentable carbohydrates and may be helpful in patients with gas-related symptoms. PMID: 17151807

Dietary supplementation with multienzyme preparations improves nutrient utilization and growth performance in weaned pigs.
Omogbenigun FO, Nyachoti CM, Slominski BA.
J Anim Sci. 2004 Apr;82(4):1053-61.

 Two experiments with young pigs (25 d of age) were conducted to investigate the effect of multienzyme preparations on nutrient digestibility, growth performance, and P utilization and excretion. In Exp. 1, 24 pigs (six pigs per treatment) were used in a 28-d performance and digestibility trial using four diets: control (no enzyme) and control supplemented with enzyme preparation A, B, or C. The control diet was formulated to meet 95% of NRC (1998) nutrient specifications (except for available P, which was at 44% NRC) and composed of corn, wheat, wheat by-products, barley, soybean meal, canola meal, and peas. All three enzyme preparations contained xylanase, glucanase, amylase, protease, invertase, and phytase activities and differed in the type of plant cell wall-degrading activities; Enzyme A contained cellulase, galactanase, and mannanase; Enzyme B contained cellulase and pectinase; and Enzyme C contained cellulase, galactanase, mannanase, and pectinase. Pigs fed enzyme-supplemented diets had higher ADG (P = 0.02) and G:F (P = 0.01) than those fed the control diet. On average, and when compared with control diet, enzyme supplementation improved (P = 0.001 to 0.04) ileal digestibility of DM (60 vs. 66%), GE (62.8 vs. 70.4%), CP (62 vs. 72%), starch (86.7 vs. 94.2%), nonstarch polysaccharides (NSP; 10.1 vs. 17.6%), and phytate (59 vs. 70%). Compared with the control, total-tract digestibility of nutrients was increased (P = 0.001 to 0.01) owing to enzyme supplementation, with Enzyme C showing the highest improvement in DM, GE, CP, starch, NSP, phytate, and P utilization. Pigs fed enzyme-supplemented diets had decreased (P = 0.04) fecal P excretion. The benefit from improved nutrient utilization with enzyme supplementation was further substantiated in a 38-d growth performance study with 48 pigs. The control and Enzyme C-supplemented diets (same as Exp. 1) were assigned to six replicate pens (four pigs per pen). The study was conducted in three phases (Phase 1 = d 0 to 7; Phase 2 = d 7 to 21; Phase 3 = d 21 to 38). Individual BW and pen feed disappearance were monitored. Average daily gain and G:F were 231 and 257 g (P = 0.01), and 0.56 and 0.63 (P = 0.001) for the control and enzyme-supplemented diets, respectively. It is evident from this study that the use of enzyme preparations may allow for cost-effective and environmentally friendly formulation of young pig diets. PMID: 15080327