Lipase Activity Research Articles

Stabilization of Eversa® Transform 2.0 lipase with sorbitol to enhance the efficiency of ultrasound-assisted biodiesel production

Ultrasound technology has emerged as a promising tool for enhancing enzymatic biodiesel production, yet the cavitation effect induced can compromise enzyme stability. This study explored the efficiency of polyols in enhancing lipase stability under ultrasound conditions to further improve biodiesel yield. The incorporation of sorbitol resulted in the highest fatty acid methyl ester (FAME) content in the ultrasound-assisted biodiesel production catalyzed by Eversa® Transform 2.0 among the investigated polyols. Furthermore, sorbitol enhanced the stability of the lipase, allowing it to tolerate up to 100 % ultrasound amplitude, compared to 60 % amplitude in its absence. Enzyme activity assays revealed that sorbitol preserved 99 % of the lipase activity, in contrast to 84 % retention observed without sorbitol under an 80 % ultrasound amplitude. Circular dichroism (CD) and fluorescence spectroscopy analyses confirmed that sorbitol enhanced lipase rigidity and preserved its conformational structure under ultrasound exposure. Furthermore, employing a stepwise methanol addition strategy in ultrasound-assisted reactions with sorbitol achieved an 81.2 wt% FAME content in 8 h with only 0.2 wt% enzyme concentration. This promising result highlights the potential of sorbitol as a stabilizing agent in ultrasound-assisted enzymatic biodiesel production, offering a viable approach for enhancing biodiesel yield and enzyme stability in industrial applications.

Enzymatic synthesis and parameters affecting on citronellyl acetate ester by trans-esterification reaction

Abstract Citronellyl acetate is a crucial component of flavor and fragrance in the food, cosmetic, and pharmaceutical sectors. In non-aqueous media, it can be successfully produced via lipase catalysis. This study focuses on the lipase-catalyzed trans-esterification of citronellol alcohol with geranyl acetate ester in a non-aqueous medium to produce citronellyl acetate. For the synthesis of citronellyl acetate, crude acetone powders isolated from several plant seedlings of black cumin, fenugreek, coriander, flax, and rape seed were examined for lipase activity. Black cumin seedling lipase had the highest level of citronella acetate production with a yield of 76.32% in 72 h of reaction time. To assess the impact of different reaction parameters on citronellyl acetate production in organic solvents, hexane was chosen as the best solvent, and black cumin seedling lipase was selected as the best biocatalyst. The highest conversion yield of ester (76.32%) was found when 0.25 M of geraniol acetate and 0.25 M of citronellol reacted at 41 oC after 72 h in the presence of 0.25 g of seedling lipase enzyme in n-hexane. It has been determined that crude black cumin seedling lipase is inexpensive yet effective and has the potential to be used industrially for the synthesis of terpene esters.

The Study of Biosurfactant Stability and The Effect on Lipase Activity

Lipase is one of hydrolase enzyme that catalyzed hydrolysis reaction of triacylglycerols into free fatty acids and monoglycerides or glycerol. These biocatalysts are widely used in several industries, namely food and pharmaceutical industry. The activity of lipase can increase significantly if the substrate forms an emulsion. Since biosurfactant has been known to have emulsification characteristic, the effect of biosurfactant addition into lipase is necessary to be investigated. It is the first report that evaluate the effect of microbial surfactant on lipase activity. The purpose of this research is to analyze the stability of biosurfactant emulsion under various conditions, such as salinity and pH as well as their potential to enhance lipase activity. Biosurfactant used was Halomonas elongata BK-AG18 from the collection of Biochemistry Research Group ITB, Bandung, Indonesia. It was found that after the addition of NaCl, there was no significant decrease in the emulsification activity of the biosurfactant. The emulsification index (IE24) of biosurfactant with several NaCl concentrations (2, 5, 10, 15, 20, and 25%) was obtained around 50%. Stability test of biosurfactant at pH range 4-10 showed the highest IE24 of biosurfactant was obtained at pH 6. The effect of biosurfactants on lipase hydrolysis activity is also discussed in this article. Lipase hydrolysis activity was tested using p-nitrophenyl palmitate substrate. The highest lipase activity was obtained after the addition of 70% biosurfactant (v/v) at 0.026 units. This study shows that biosurfactant from H. elongata BK-AG18 has the potential to increase lipase activity.

Potassium diformate affects the growth and development of broilers by improving intestinal function and digestive enzyme activity

Gut health of broiler chickens is essential for production performance. The present study aimed to evaluate the impact of dietary supplementation with potassium diformate (KDF) on growth performance and intestinal health in broiler chickens. A total of 180 Arbor Acres (AA) broiler chickens were randomly allocated into 3 treatments, with 6 replicates, containing 10 chicks in each replicate. The treatment groups were: control group (CON) was fed a basal diet; KDF-4 groups fed the basal diet with 4 g/kg KDF; KDF-8 groups fed the basal diet with 8 g/kg KDF. The experiment period lasted for 42 d. During the starter phase, the ADFI and F/G of broilers in KDF groups were lower (P < 0.05) compared to the CON group. Furthermore, the BW and ADG in KDF-4 group was improved (P<0.05). The treatment groups exhibited a significant increase (P < 0.05) in both ADG and ADFI during the grower and overall phase. Moreover, the F/G in KDF-4 group was lower (P < 0.05) compared to the CON and KDF-8 groups. The semi-eviscerated weight rate (SEWR), eviscerated carcass weight rate (ECWR), pectoral muscle rate (PMR), and leg muscle rate (LMR) of broilers were improved (P < 0.05) in KDF groups. The serum levels of glucose (GLU) and UREA (UA) were significantly higher (P < 0.05) in KDF-8 group. Additionally, the nutrient apparent utilization rate of dry matter (DM), energy (EE), and crude protein (CP) were improved (P < 0.05) in KDF-4 group. The villus height (VH) and villus height to crypt depth ratio (V/C) of duodenum, jejunum, and ileum were higher (P < 0.05) in KDF groups compared to the CON group, while crypt depth (CD) was significantly reduced (P < 0.05). The digestive enzyme activities of lipase (LIP), amylase (AMS), or trypsin (TPS) were significantly enhanced (P < 0.05) in the intestinal chyme, while the total bacterial count, Escherichia coli, Lactobacilli, Bifidobacteria, and Bacillus were reduced (P < 0.05) in the ileum. This study demonstrates that the inclusion of KDF in the diet of broilers leads to improvements in growth, slaughter performance, nutrient utilization rate, and maintenance of intestinal health.

Dietary supplementation of protease and organic acid in poultry by-product meal-based diet in broilers.

This study investigated the impact of supplementation of protease and organic acid on growth performance and other biological parameters in broilers fed poultry byproduct meal (PBM) based diet. Five hundred 1-day-old broiler chicks (Ross 308) were distributed into five treatments with 5 replicates, each pen having 20 birds, and fed each group one of five isocaloric and isonitrogenous diets in two phases: stater phase (1 to 21 days) metabolizable energy (ME) 3000 kcal/kg; crude protein (CP) 22%, and a finisher phase (22 to 35 days) ME 3,200 kcal/kg; CP 19.5%. The dietary treatments were: i) standard broiler ration (Cont); ii) The control diet with 25% of the soybean meal replaced by PBM on an equivalent protein basis (PBM); iii) PBM diet supplemented with 0.5 g/kg of protease (PBMP); iv) PBM diet supplemented with 1 g/kg organic acid (PBMO); and v) PBM diet addition with 0.5 g/kg protease and 1 g/kg organic acid (PBMPO). The overall data showed that feed conversion ratio was improved (p<0.05) in the PBMP group. Apparent CP digestibility was higher (p<0.05) in both Cont and PBMP groups. Jejunal villus height increased (p<0.05) in PBMP and PBMPO groups, while only the PBMO group exhibited a higher (p<0.05) crypt depth. Lipase activity was increased (p<0.05) in the PBMP, PBMO, and PBMPO dietary treatments. However, trypsin activity showed a significant increase (p<0.05) in the PBMP and PBMO groups. Serum biochemistry increased (p<0.05) globulin and total protein levels in the PBMP group. PBM could partially replace the soybean meal with supplementation of either protease or organic acid in broiler diets without impairing overall growth performance. Furthermore, careful optimization must be considered when combining protease and organic acids.

Designing biodegradable aqueous biphasic systems for the selective separation of enzymes

In this work, the salting-out strength of reline eutectic mixture in aqueous solution of the non-ionic surfactant Triton X-102 has been demonstrated at temperatures between 298.15 and 323.15 K as a preliminary step to be employed in downstream operations of a biotechnological process. The phase diagrams data of reline-based Aqueous Two Phase Systems (ATPS) have been correlated using empirical models based on exponential and polynomial equations. The tie-lines have been empirically ascertained through physical properties characterization and Othmer-Tobias equation was proposed to correlate those data. After having studied the effect of the proposed reline eutectic mixture on two model enzymes (protease and lipase), the performance of this ATPS for selectively separating them was evaluated for three different feed compositions on the same tie-line, demonstrating their pertinence to segregate the lipase to one of the phases (yield about 95 %) and the protease to the other phase (yield about 98 %). These promising results are interesting for application in biotechnological reactions where proteases have been proved to be deleterious for lipases activity, as the biocompatibility of the proposed platform does not involve drastic negative effects on any of the enzymes.

Effects of oxidized fish oil diet supplemented with tea polyphenols on intestinal health and liver metabolism of spotted sea bass (Lateolabrax maculatus)

In this study, the effects of oxidized fish oil diet supplemented with tea polyphenols (TP) on the intestinal health and liver metabolism of spotted sea bass (Lateolabrax maculatus) were investigated. Five kinds of diets were designed. They were the negative control (NC) group without antioxidants and TP, the positive control (PC) group with antioxidants, three experimental groups were supplemented with 0.05 %, 0.10 %, and 0.15 % TP (TP1, TP2, and TP3). The fish with a mean body weight of 11.43±0.02 g was fed on five diets for 56 days. The results showed that TP1 improved lipase (LPS) activity. The intestinal superoxide dismutase (SOD) activity of TP1 was significantly higher than that of the NC group and was not different from that of the PC group. Histological observation of the intestine showed that the height of the intestinal villus was significantly increased and the width of the intestinal villus was significantly decreased in TP1, while the muscular thickness layer did not change significantly. Furthermore, inflammatory symptoms were present in the intestines of all groups, with improvements observed in TP1 and TP3. By intestinal microbial analysis, the results showed that the ACE and Chao1 indexes of TP1 were significantly higher than those of the NC and PC groups, and the Shannon and Simpson indexes were significantly lower than those of the NC and PC groups. Regarding the structure of the microbiome community, there were a lot more Fusobacteriota in TP2 and TP3 at the phylum level, and there were a lot more Cetobacterium at the genus level. The metabolomic analysis of TP1's liver showed that it controls the metabolism of amino acids, nucleotides, cofactors, vitamins, energy, and carbohydrates. The results showed that adding TP to the oxidized fish oil diet enhanced the intestinal health and liver metabolism of spotted sea bass.

Effects of Yacon (Smallanthus sonchifolius) Juice Byproduct Administered Using Different Feeding Methods on the Growth Performance, Digestive Enzyme Activity, Antioxidant Status, and Disease Resistance against Streptococcus iniae of Juvenile Black Rockfish (Sebastes schlegelii)

This study was designed to evaluate the effects of yacon (Smallanthus sonchifolius) juice byproduct (YJB) on the growth performance, digestive and antioxidant enzyme activities, and disease resistance against Streptococcus iniae of juvenile black rockfish (Sebastes schlegelii) based on different feeding schedules. Four different YJB feeding strategies were evaluated: feeding the fish a basal diet continuously (control, T0), feeding them YJB (2.5 g/kg) continuously (T1), feeding them YJB for 1 day and the basal diet the next day (T2), and feeding them YJB for 1 day and the basal diet for the following 2 days (T3). No difference in survival among the treatments was found after the 8-week feeding trial (p &gt; 0.05). However, the T1 and T2 groups exhibited significant enhancements in final body weight, weight gain, and specific growth rate compared with the T0 and T3 groups. Furthermore, the T1 and T2 groups showed a significant improvement in feed consumption, feed efficiency, and protein efficiency ratio compared with the T0 and T3 groups. No significant differences in the condition factor or viscerosomatic and hepatosomatic indices were observed among all the groups. Intestinal amylase, trypsin, and lipase activity was significantly (p &lt; 0.05) higher in the T1 and T2 groups than in the T0 and T3 groups. Lysozyme, superoxide dismutase, and catalase activity along with glutathione peroxidase content were significantly (p &lt; 0.05) higher under all YJB feeding regimens than those under the control treatment. The survival rates in all the YJB treatment groups after the S. iniae challenge were significantly (p &lt; 0.05) enhanced. In conclusion, we recommend offering YJB at day-to-day intervals to improve growth performance, digestive enzyme activity, antioxidant status, and disease resistance against S. iniae.

Marine copepod culture as a potential source of bioplastic-degrading microbiome: The case of poly(butylene succinate-co-adipate)

The poly(butylene succinate-co-adipate) (PBSA) is emerging as environmentally sustainable polyester for applications in marine environment. In this work the capacity of microbiome associated with marine plankton culture to degrade PBSA, was tested. A taxonomic and functional characterization of the microbiome associated with the copepod Acartia tonsa, reared in controlled conditions, was analysed by 16S rDNA metabarcoding, in newly-formed adult stages and after 7 d of incubation. A predictive functional metagenomic profile was inferred for hydrolytic activities involved in bioplastic degradation with a particular focus on PBSA. The copepod-microbiome was also characterized in newly-formed carcasses of A. tonsa, and after 7 and 33 d of incubation in the plankton culture medium. Copepod-microbiome showed hydrolytic activities at all developmental stages of the alive copepods and their carcasses, however, the evenness of the hydrolytic bacterial community significantly increased with the time of incubation in carcasses. Microbial genera, never described in association with copepods: Devosia, Kordia, Lentibacter, Methylotenera, Rheinheimera, Marinagarivorans, Paraglaciecola, Pseudophaeobacter, Gaiella, Streptomyces and Kribbella sps., were retrieved. Kribbella sp. showed carboxylesterase activity and Streptomyces sp. showed carboxylesterase, triacylglycerol lipase and cutinase activities, that might be involved in PBSA degradation. A culturomic approach, adopted to isolate bacterial specimen from carcasses, led to the isolation of the bacterial strain, Vibrio sp. 01 tested for the capacity to promote the hydrolysis of the ester bonds. Granules of PBSA, incubated 82 d at 20 °C with Vibrio sp. 01, were characterized by scanning electron microscopy, infrared spectroscopy, thermogravimetric analysis, and differential scanning calorimetry, showing fractures compared to the control sample, and hydrolysis of ester bonds. These preliminary results are encouraging for further investigation on the ability of the microbiome associated with plankton to biodegrade polyesters, such as PBSA, and increasing knowledge on microorganisms involved in bioplastic degradation in marine environment.

Effects of dietary marine red yeast supplementation on growth performance, antioxidant, immunity, lipid metabolism and mTOR signaling pathway in Juvenile Tilapia (Oreochromis niloticus)

A 60-day feeding trial was conducted to investigate the effects of dietary marine red yeast supplementation on growth performance, body composition, antioxidant, immunity, lipid metabolism, and mTOR signaling pathway in juvenile GIFT tilapia (Initial weight 21.12 ± 0.86 g). Five isonitrogenous and isolipidic experimental diets were formulated with graded levels of marine red yeast: 0 % (control group), 0.25 %, 0.50 %, 0.75 %, and 1.00 %. The results showed that different marine red yeast levels significantly increased (p < 0.05) the final weight, weight gain rate, specific growth rate, daily growth index, protein efficiency ratio, and condition factor, while significantly decreased (p < 0.05) the feed conversion ratio, viscera index, and hepatosomatic index. Different marine red yeast levels significantly increased (p < 0.05) the activities of foregut lipase, hindgut lipase, foregut α-amylase, midgut α-amylase, hindgut α-amylase, superoxide dismutase, catalase, glutathione peroxidase, and total antioxidant capacity, while the malondialdehyde content expressed an opposite trend. Different marine red yeast levels significantly increased (p < 0.05) the activities (contents) of lysozyme, alkaline phosphatase, immunoglobulin M, and complement protein 3 in serum. Different marine red yeast levels significantly increased (p < 0.05) the relative expression of phosphoryl inositol-3-kinase, protein kinase B, mammalian target of rapamycin, ribosome S6 protein kinase 1, cholesterol response element binding protein 1, peroxisome proliferator-activated receptor γ, leptin, acetyl-CoA carboxylase α, fatty acid synthase, and glucose-6-phosphate dehydrogenase in liver, while the relative expression of PPARα, carnitine palmitoyl transferase 1, lipoprotein lipase, and hormone sensitive lipase expressed an opposite trend. In conclusion, dietary supplementation with marine red yeast significantly increased the growth performance, antioxidant capacity, initial immunity, lipid synthesis, and mTOR signaling pathway in juvenile GIFT tilapia. Based on the second-order polynomial regression analysis, the optimal dietary marine red yeast level was between 0.53 % and 0.60 %.

Hydrolytic enzyme activity in high-rate anaerobic reactors treating municipal wastewater in temperate climates

Particulate matter hydrolysis is the bottleneck in anaerobic treatment of municipal wastewater in temperate climates. Low temperatures theoretically slow enzyme-substrate interactions, hindering utilization kinetics, but this remains poorly understood. β-glucosidase, protease, and lipase activities were evaluated in two pilot-scale upflow anaerobic sludge blanket (UASB) reactors, inoculated with different sludges and later converted to anaerobic membrane bioreactors (AnMBRs). Despite similar methane production and solids hydrolysis rates, significant differences emerged. Specific activity peaked at 37 °C, excluding the predominance of psychrophilic enzymes. Nevertheless, the Michaelis-Menten constant (Km) indicated high enzyme-substrate affinity at the operational temperature of 15–20 °C, notably greater in AnMBRs. It is shown, for the first time, that different seed sludges can equally adapt, as hydrolytic enzymatic affinity to the substrate reached similar values in the two reactors at the operational temperature and identified that membrane ultrafiltration impacted hydrolysis by a favourable enzyme Michaelis-Menten constant.

Targeted Screening of Curcumin Derivatives as Pancreatic Lipase Inhibitors Using Computer-Aided Drug Design.

Curcumin has demonstrated promising preclinical antiobesity effects, but its low bioavailability makes it difficult to exert its full effect at a suitable dose. The objective of this study was to screen curcumin derivatives with enhanced bioavailability and lipid-lowering activity under the guidance of computer-aided drug design (CADD). CAAD was used to perform virtual assays on curcumin derivatives to assess their pharmacokinetic properties and effects on pancreatic lipase activity. Subsequently, 19 curcumin derivatives containing 5 skeletons were synthesized to confirm the above virtual assay. The in vitro pancreatic lipase inhibition assay was employed to determine the half-maximal inhibitory concentration (IC50) of these 19 curcumin derivatives. Based on CADD analysis and in vitro pancreatic lipase inhibition, 2 curcumin derivatives outperformed curcumin in both aspects. Microscale thermophoresis (MST) experiments were employed to assess the binding equilibrium constants (K d) of the aforementioned 2 curcumin derivatives, curcumin, and the positive control drug with pancreatic lipase. Through virtual screening utilizing a chemoinformatics database and molecular docking, 6 derivatives of curcumin demonstrated superior solubility, absorption, and pancreatic lipase inhibitory activity compared to curcumin. The IC50 value for 1,7-bis(4-hydroxyphenyl)heptane-3,5-dione (C4), which displayed the most effective inhibitory effect, was 42.83 μM, while the IC50 value for 1,7-bis(4-hydroxy-3-methoxyphenyl)heptane-3,5-dione (C6) was 98.62 μM. On the other hand, the IC50 value for curcumin was 142.24 μM. The MST experiment results indicated that the K d values of C4, C6, and curcumin were 2.91, 18.20, and 23.53 μM, respectively. The results of the activity assays exhibited a relatively high degree of concordance with the outcomes yielded by CADD screening. Under the guidance of CADD, the targeted screening of curcumin derivatives with excellent properties in this study exhibited high-efficiency and low-cost benefits.

Effects of Olive Oil and Tween 80 on Production of Lipase by Yarrowia Yeast Strains

Lipase is one of the most commonly used biocatalysts in the food, pharmaceutical and cosmetic industries, and can be produced by Yarrowia lipolytica yeast. Despite the intensive studies of lipase from Yarrowia, there are still many open questions regarding the enzyme secretion process, especially by new isolates of this genus as well as the effect of substrates or surfactants, or both on the production of lipase. This research focused on the effect of olive oil and surfactant Tween 80 including the optimisation of the concentration of these compounds on the production of lipase by some novel Yarrowia isolates. Moreover, the optimal environmental parameters (pH, temperature) of crude enzyme synthetised by Yarrowia strains were determined. All investigated strains were able to produce lipase in both intracellular and extracellular fractions. The extracellular lipase activities were higher than the intracellular ones (Y. divulgata Y.02062 and Yarrowia lipolytica 854/4 147 U/L, 80 U/L and 474 U/L, 122 U/L, respectively). In the case of extracellular lipase, supplementing olive oil and Tween 80 enhanced significantly the synthesis and secretion of the enzyme. The lipase activity can even be enhanced by 20 times higher from 25 U/L to 474 U/L in the case of Yarrowia lipolytica 854/4 strain. In the case of intracellular, supplementation of Tween 80 generally reduces lipase activity except for the Y. lipolytica 1/4 strain, which was affected by two times the increase. The optimised concentration of olive oil and Tween 80 were determined for Y. divulgata Y.02062, Y. divulgata 5257, Y. lipolytica 1/4, and Yarrowia lipolytica 854/4 strains as 1.6% olive oil and 0.09% Tween 80, 1.6% olive oil and 0.06% Tween 80, 1.4% olive oil and 0.09% Tween 80 as well as 1.6% olive oil and 0.065% Tween 80, respectively. The optimum pH and temperature of crude lipases (intra and extracellular) synthetised by the tested Yarrowia lipolytica and Y. divulgata yeast strains were found to be pH 7.2 and 37 °C, respectively. Our results confirmed that the new isolate Y. divulgata is a very promising species for further development for industrial use as Y. lipolytica.

Effects of florfenicol on the midgut physiological function of Bombyx mori, based on the diversity of intestinal microbiota

AbstractFlorfenicol (FF) is a new antibiotic commonly used in sericulture. This study aimed to examine the effects of low (LC: 0.06 g/L), medium (MC: 0.12 g/L), and high (HC: 1.2 g/L) concentrations of FF on the midgut physiological functions of the silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae). The results showed that the body weight and the whole cocoon weight of silkworms decreased and the development duration of the fifth instar was prolonged in the HC group. The structure of intestinal microbiota of silkworm larvae was changed by high FF exposure. Specifically, the abundance of conditional pathogens (Curtobacterium, Sanguibater, etc.) was significantly increased, whereas the abundance of Pseudomonas and Pedobacter was decreased significantly. Additionally, the intestinal reactive oxygen species level at 72 h was significantly elevated, and the muscle layer had become loose, together with the appearance of gut goblet cell atrophy. We observed that the increase in Lactobacillus abundance led to a reduction in intestinal fluid pH, resulting in decreased α‐amylase and protease activities, whereas lipase activity exhibited an elevation. The results demonstrated that the accumulation of peroxides induced by exposure to 1.2 g/L FF caused intestinal damage, and the decrease in pH resulting from alterations in microbial composition and structure affected digestive enzyme activity, collectively leading to physiological impairment of the silkworm midgut. These findings provide a valuable reference for the safe use of FF in sericulture.

Evaluation of soybean meal replacement by Clostridium autoethanogenum protein in diets for juvenile grass carp (Ctenopharyngodon idellus)

An investigation into the possible substitution of Clostridium autoethanogenum protein (CAP) for soybean meal (SBM) in the diet of grass carp (Ctenopharyngodon idellus) was carried out. During a 56-day feeding trial, 750 grass carp were fed five experimental diets with different levels of CAP: SBM (control group), CAP25, CAP50, CAP75, and CAP100, corresponding to dietary SBM replacements levels of 0%, 25%, 50%, 75%, and 100%. The results showed that dietary CAP significantly increased FW, WGR, and PER in CAP25 group (P < 0.05), but complete substitution showed the opposite trend (P < 0.05). The liver antioxidant capacity was improved in CAP25 group manifested by the enhanced SOD, GPx and CAT activities, reduced ROS, and MDA contents, and the up-regulated cuznsod, mnsod, gpx1, gpx4, gsto and gr transcript levels mediated by Keap1-Nrf2 signaling pathway (P < 0.05). The intestinal inflammation was relieved in CAP25 group evidenced by the down-regulated il-β, il-6 and tnf-α expressive abundance through NF-κB pathway (P < 0.05). Additionally, the intestinal mucosal barrier function was enhanced by the upregulation of tight junction proteins (occludin, zo-1 and zo-2) in the CAP25 group. Whereas the intestinal folds showed varying degrees of breakage in CAP100 group. The increased amylase and lipase activities, intestinal folds height, and goblet cell numbers were found in CAP25 group (P < 0.05). The gut microbiota revealed no significant differences in the alpha diversity indices across treatments (P > 0.05). Besides, the abundance of Bacteroides in the CAP25 group was increased, while the abundance of Erysipelatoclosridium in the CAP100 group was increased. Summarily, CAP could replace 25% of SBM with positive influences on growth, antioxidant capacity, immunity and intestinal microbiota for juvenile grass carp. Conversely, high levels of CAP impaired intestinal physical barrier and caused intestinal microbiota disturbance, resulting in negatively affect the growth performance.

Effects of supplemental Glycyrrhiza polysaccharide on growth performance and intestinal health in weaned piglets

In this study, we investigated the effects of supplemental Glycyrrhiza polysaccharide (GCP) on growth performance and intestinal health of weaned piglets. Ninety piglets weaned at 28 days of age were randomly allocated to three groups with five replicates per treatment. Piglets were fed the following diets for 28 days: (1) CON (control group), basal diet; (2) G500, CON + 500 mg/kg GCP; (3) G1000, CON + 1000 mg/kg GCP. The results showed that supplementation with 1000 mg/kg GCP increased the average daily gain (ADG) and decreased the feed-to-gain ratio (F/G) (P < 0.05). Serum diamine oxidase (DAO) and D-lactic acid (DL-A) levels were lower in the G1000 group (P < 0.05). Dietary GCP 1000 mg/kg improved mucosal trypsin activity in the duodenum, jejunum and ileum and increased lipase and amylase activity in the jejunum (P < 0.05). Moreover, in the G1000 group, ZO-1, claudin 1 and occludin levels were increased in the jejunum mucosa, whereas interleukin-1β (IL-1β) and IL-6 levels were decreased (P < 0.05). The 16S rRNA gene analysis indicated that dietary 1000 mg/kg GCP altered the jejunal microbial community, with increased relative abundances of beneficial bacteria. In conclusion, dietary GCP 1000 mg/kg can improve growth performance, digestive enzyme activity, intestinal immunity, barrier function and microbial community in weaned piglets.

Comparative studies on the intestinal health of wild and cultured ricefield eel (Monopterus albus).

Fish intestinal health under intensive aquaculture mode plays an important role in growth, development, and immune function. The present study was aimed to systematically investigate the differences of intestinal health between wild and cultured Monopterus albus by biochemical parameters, histomorphology, and molecular biology. A total of 15 healthy M. albus per group, with an average body weight of 45g, were sampled to analyze intestinal health parameters. Compared with wild fish, the cultured M. albus in the foregut had lower trypsin, lipase, SOD, CAT, T-AOC, and GSH-Px activities (P < 0.05) and higher amylase activity and MDA content (P < 0.05). The villus circumference and goblet cells in the cultured group were significantly lower than those in the wild group (P < 0.05). In addition, the cultured fish showed lower relative expression levels of occludin, zo-1, zo-2, claudin-12, claudin-15, mucin5, mucin15, lysozyme, complement 3, il-10, tgf-β1, tgf-β2, and tgf-β3 (P < 0.05) and higher il-1β, il-6, il-8, tnf-a, and ifnγ mRNA expressions than those of wild fish (P < 0.05). In terms of gut microbiota, the cultured group at the phylum level displayed higher percentages of Chlamydiae and Spirochaetes and lower percentages of Firmicutes, Bacteroidetes, Actinobacteria, Cyanobacteria, and Verrucomicrobia compared to the wild group (P < 0.05). At the genus level, higher abundances of Pseudomonadaceae_Pseudomonas and Spironema and lower abundances of Lactococcus and Cetobacterium were observed in the cultured group than in the wild group (P < 0.05). To our knowledge, this is the first investigation of the intestinal health status between wild and cultured M. albus in terms of biochemistry, histology, and molecular biology levels. Overall, the present study showed significant differences in intestinal health between wild and cultured M. albus and the main manifestations that wild M. albus had higher intestinal digestion, antioxidant capacity, and intestinal barrier functions than cultured M. albus. These results would provide theoretical basis for the subsequent upgrading of healthy aquaculture technology and nutrient regulation of intestinal health of cultured M. albus.

Construction of Pickering emulsion and its application in enzymatic synthesis of phytosteryl oleate

In this study, Janus nanoparticles were prepared and used to construct Pickering emulsion as an emulsifier in presence of lipase. The results showed that Janus nanoparticles could be used to stable water phase in emulsion and the average particle size of emulsion was 60 μm. Phytosterol ester was successfully synthesized in Pickering emulsion catalytic system by using oleic acid as acyl accepter, and the phytosterol conversion reached 91.65 ± 2.07% under the following conditions: emulsifier amount of 1.0 wt%, oil-water ratio of 6:1, phytosterol concentration of 110 mmol/L, lipase dosage of 20 g/L, reaction temperature of 45 °C. Compared with biphasic catalytic system, Pickering emulsion catalytic system could enhance the catalytic efficiency and shorten the reaction time due to large interface area and short molecule diffusion distance. At the end of reaction, Janus nanoparticles could be recovered by simple centrifugation, the relative activity of lipase remained 72.89 ± 1.83% after repeated use for 6 times. Overall, this paper provides a novel and high efficiency interfacial biocatalysis technique for the synthesis of phytosteryl oleate.

The American cockroach (Periplaneta americana) residue could partially replace the dietary puffed soybean meal in the Three-yellow chickens

Periplaneta americana residue is a byproduct of using Periplaneta americana in pharmaceutical research and development for extracting active ingredients. Three hundred Three-yellow chickens were selected for the experiment and randomly divided into 6 groups (5 replications per group, 10 chickens per replicate): the control group (group A) was fed a basal ration, and the experimental groups (groups B, C, D, E, and F) were fed experimental diets in which P. americana residue replaced puffed soybean meal at proportions of 20, 40, 60, 80, and 100%, respectively, for a period of 42 d. The aim was to assess the impact of different levels of P. americana residue on the growth, survival, intestinal morphology, digestive enzyme activity, intestinal flora, and intestinal transcriptional responses of Three-yellow chickens. The results indicated that the increase in P. americana residue levels had a linear and quadratic impact on the average daily gain (ADG) and feed conversion ratio (FCR), respectively. The ADG was notably greater in the 40% group than in the 100% group, while the FCR was significantly lower in the 20% and 40% groups than in the 100% group (P < 0.05). Protease, lipase, and amylase activities exhibited a quadratic increase with increasing concentrations of P. americana residue (P < 0.05). Protease and lipase activities were notably greater in the 20% and 40% groups than in the 0% group (control group), amylase activity was significantly greater in the 40% group than in the 0% group (control group) (P < 0.05). Duodenal crypt depth (CD) decreased quadratically with increasing P. americana residue (P < 0.05). The duodenal villus height/crypt depth ratio (V/C) was significantly lower in the 100% group than in the 60% group (P < 0.05). The intestinal villus height (VH) increased quadratically with increasing levels of P. americana residue. The VH in the 60% group was significantly greater than that in the 0% (control group), 20, 80, and 100% groups (P < 0.05). The Chao and Ace indices demonstrated linear and quadratic increases with increasing levels of P. americana residue, while the Pd index showed a quadratic increase with increasing levels of P. americana residue (P < 0.05). The relative abundance profile of Lactobacillus exhibited a linear and quadratic decrease with increasing levels of P. americana residue, with the 100% group showing a significantly lower abundance than the 0% (control group) and 40% groups (P < 0.05). The transcriptome results showed that P. americana residue could enhance the digestive system by promoting vitamin, fat, carbohydrate digestion and absorption, cholesterol metabolism, etc. In conclusion, P. americana residue can replace puffed soybean meal without negatively affecting the growth performance of three-yellow chickens. The low and medium groups had positive effects on the growth performance, digestive enzyme activity, intestinal morphology, intestinal flora, and substance digestion and absorption of three-yellow chickens. The recommended replacement of P. americana residue for puffed soybean meal in the diets of three-yellow chickens ranged from 20% to 60%.

Assessment of Fungal Lytic Enzymatic Extracts Produced Under Submerged Fermentation as Enhancers of Entomopathogens’ Biological Activity

The application of enzymes in agricultural fields has been little explored. One potential application of fungal lytic enzymes (chitinases, lipases, and proteases) is as an additive to current biopesticides to increase their efficacy and reduce the time of mortality. For this, a screening of lytic overproducer fungi under submerged fermentation with a chemical-defined medium was performed. Then, the enzymatic crude extract (ECE) was concentrated and partially characterized. This characterization consisted of measuring the enzymatic activity (lipase, protease and, chitinase) and determining the enzyme stability after storage at temperatures of − 80, − 20 and, 4 °C. And lastly, the application of these concentrated enzymatic crude extracts (C-ECE) as an enhancer of spores-based fungal biopesticide was proven. Beauveria were not as good producers of lytic enzymes as the strains from Trichoderma and Metarhizium. The isolate M. robertsii Mt015 was selected for the co-production of chitinases and proteases; and the isolate T. harzianum Th180 for co-production of chitinases, lipases, and proteases. The C-ECE of Mt015 had a protease activity of 18.6 ± 1.1 U ml−1, chitinase activity of 0.28 ± 0.01 U ml−1, and no lipase activity. Meanwhile, the C-ECE of Th180 reached a chitinase activity of 0.75 U ml−1, lipase activity of 0.32 U ml−1, and protease activity of 0.24 U ml−1. Finally, an enhancing effect of the enzymatic extracts of M. robertsii (66.7%) and T. harzianum (43.5%) on the efficacy of B. bassiana Bv064 against Diatraea saccharalis larvae was observed. This work demonstrates the non-species-specific enhancing effect of enzymatic extracts on the insecticidal activity of conidial-based biopesticides, which constitutes a contribution to the improvement of biological control agents’ performance.