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1.
Antioxidants (Basel) ; 13(5)2024 May 05.
Artículo en Inglés | MEDLINE | ID: mdl-38790674

RESUMEN

Soybean meal (SBM) is a prevailing plant protein supplement in animal diets because of its nutritional value and availability. This review paper explores the significance of SBM and processed soy products, emphasizing their nutritional and bioactive components, such as isoflavones and soyasaponins. These compounds are known for their antioxidant and anti-inflammatory properties and are associated with a reduced prevalence of chronic diseases. However, the presence of antinutritional compounds in SBM presents a significant challenge. The paper evaluates various processing methods, including ethanol/acid wash, enzyme treatment, and fermentation, which are aimed at enhancing the nutritional value of soy products. It highlights the significance to maintain a balance between nutritional enhancement and the preservation of beneficial bioactive compounds, emphasizing the importance of different processing techniques to fully exploit the health benefits of soy-based products. Therefore, this review illuminates the complex balance between nutritional improvement, bioactive compound preservation, and the overall health implications of soy products.

2.
Anim Nutr ; 14: 235-248, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37600837

RESUMEN

This study investigated the effects of using soy protein concentrate (SPC) to replace animal protein supplements on mucosa-associated microbiota, intestinal health, and growth performance of nursery pigs. Fifty-six newly weaned pigs (BW = 6.4 ± 0.6 kg) were allotted to 5 treatments in a randomized complete block design. Pigs were fed for 35 d in 3 phases (P; 1, 2, 3) for 10, 12, 13 d, respectively. Dietary treatments were: (1) basal diet with fish meal (P1: 4%, P2: 2%, and P3: 1%), poultry meal (P1: 10%, P2: 8%, and P3: 4%), and blood plasma (P1: 4%, P2: 2%, and P3: 1%), where SPC replacing none (NC); (2) basal diet with SPC replacing fish meal (RFM); (3) basal diet with SPC replacing poultry meal (RPM); (4) basal diet with SPC replacing blood plasma (RBP); and (5) basal diet with SPC replacing all animal protein supplements (PC). Growth performance was recorded for each phase. Pigs were euthanized on d 35 to collect jejunal mucosa and tissue to evaluate intestinal health and microbiota, and ileal digesta to measure apparent ileal digestibility (AID) of nutrients. Data were analyzed using the MIXED procedure of SAS. Overall, RFM, RPM, and RBP did not affect growth performance, whereas PC decreased (P < 0.05) ADG and ADFI. The RPM increased (P < 0.05) Prevotella stercorea and decreased (P < 0.05) Helicobacter rappini. The PC decreased (P < 0.05) H. rappini, whilst increasing (P < 0.05) Prevotella copri, Propionibacterium acnes, and Pelomonas aquatica. The RFM tended to increase (P = 0.096) immunoglobulin A in the jejunum. The PC tended to decrease (P = 0.078) jejunal crypt cell proliferation. There were no differences in the villus height, AID of nutrients, intestinal inflammation, and intestinal oxidative stress among treatments. In conclusion, SPC can replace fish meal, poultry meal, or blood plasma individually without affecting growth performance and intestinal health, and AID of nutrients of nursery pigs. Particularly SPC replacing poultry meal benefitted intestinal health by reducing H. rappini and increasing P. stercorea. However, SPC replacing all three animal protein supplements reduced growth of nursery pigs mainly by reducing feed intake.

3.
Toxins (Basel) ; 15(7)2023 06 30.
Artículo en Inglés | MEDLINE | ID: mdl-37505702

RESUMEN

This study aimed to investigate the efficacy of a feed additive containing bentonite and enzymatically hydrolyzed yeast on the intestinal health and growth of newly weaned pigs under chronic dietary exposure to fumonisin and aflatoxin. Newly weaned pigs were randomly allotted to one of four possible treatments: a control diet of conventional corn; a diet of corn contaminated with fumonisin and aflatoxin; a diet of mycotoxin-contaminated corn with 0.2% of feed additive; and a diet of mycotoxin contaminated corn with 0.4% of feed additive. We observed lower average weight gain and average daily feed intake in pigs that were fed only mycotoxin-contaminated corn compared to the control group. Feed additive supplementation linearly increased both average weight gain and feed intake, as well as tumor necrosis factor-alpha. In the jejunum, there was an observed decrease in immunoglobulin A and an increase in claudin-1. Additionally, feed additive supplementation increased the villus height to crypt depth ratio compared to the control. In conclusion, feed additives containing bentonite and enzymatically hydrolyzed yeast could mitigate the detrimental effects of mycotoxins on the growth performance of newly weaned pigs by improving intestinal integrity and positively modulating immune response.


Asunto(s)
Aflatoxinas , Fumonisinas , Micotoxinas , Porcinos , Animales , Fumonisinas/toxicidad , Saccharomyces cerevisiae , Bentonita , Aflatoxinas/toxicidad , Suplementos Dietéticos/análisis , Dieta/veterinaria , Micotoxinas/toxicidad , Aumento de Peso , Alimentación Animal/análisis
4.
J Anim Sci Biotechnol ; 14(1): 89, 2023 Jul 02.
Artículo en Inglés | MEDLINE | ID: mdl-37393326

RESUMEN

BACKGROUND: Soy protein supplements, with high crude protein and less antinutritional factors, are produced from soybean meal by different processes. This study evaluated the comparative effects of various soy protein supplements replacing animal protein supplements in feeds on the intestinal immune status, intestinal oxidative stress, mucosa-associated microbiota, and growth performance of nursery pigs. METHODS: Sixty nursery pigs (6.6 ± 0.5 kg BW) were allotted to five treatments in a randomized complete block design with initial BW and sex as blocks. Pigs were fed for 39 d in 3 phases (P1, P2, and P3). Treatments were: Control (CON), basal diet with fish meal 4%, 2%, and 1%, poultry meal 10%, 8%, and 4%, and blood plasma 4%, 2%, and 1% for P1, P2, and P3, respectively; basal diet with soy protein concentrate (SPC), enzyme-treated soybean meal (ESB), fermented soybean meal with Lactobacillus (FSBL), and fermented soybean meal with Bacillus (FSBB), replacing 1/3, 2/3, and 3/3 of animal protein supplements for P1, P2, and P3, respectively. Data were analyzed using the MIXED procedure in SAS 9.4. RESULTS: The SPC did not affect the BW, ADG, and G:F, whereas it tended to reduce (P = 0.094) the ADFI and tended to increase (P = 0.091) crypt cell proliferation. The ESM did not affect BW, ADG, ADFI, and G:F, whereas tended to decrease (P = 0.098) protein carbonyl in jejunal mucosa. The FSBL decreased (P < 0.05) BW and ADG, increased (P < 0.05) TNF-α, and Klebsiella and tended to increase MDA (P = 0.065) and IgG (P = 0.089) in jejunal mucosa. The FSBB tended to increase (P = 0.073) TNF-α, increased (P < 0.05) Clostridium and decreased (P < 0.05) Achromobacter and alpha diversity of microbiota in jejunal mucosa. CONCLUSIONS: Soy protein concentrate, enzyme-treated soybean meal, and fermented soybean meal with Bacillus could reduce the use of animal protein supplements up to 33% until 7 kg body weight, up to 67% from 7 to 11 kg body weight, and entirely from 11 kg body weight without affecting the intestinal health and the growth performance of nursery pigs. Fermented soybean meal with Lactobacillus, however, increased the immune reaction and oxidative stress in the intestine consequently reducing the growth performance.

5.
J Anim Sci ; 100(10)2022 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-35950990

RESUMEN

This study was to evaluate the effects of soy protein concentrate (SPC) supplementation replacing animal protein supplements on intestinal immune status, intestinal oxidative stress status, nutrient digestibility, mucosa-associated microbiota, and growth performance of nursery pigs. Thirty-two newly weaned pigs at 21 d of age with 6.4 ± 0.4 kg body weight (BW) were allotted to four treatments in a randomized complete block design with initial BW and sex as blocks. Pigs were fed for 35 d in three phases. Dietary treatments were SPC 0% (diets with fish meal 4/2/1%, poultry meal 10/8/4%, blood plasma 4/2/1%, and crude protein 24.6/22.6/20.9% for phase 1/2/3, respectively), SPC 33%, SPC 66%, and SPC 100% (SPC 0% diets with SPC replacing 33/66/100% of animal protein supplements, respectively). Pigs were euthanized on day 35 to collect jejunal mucosa and tissues to evaluate intestinal immune status, intestinal oxidative stress status, intestinal morphology, and mucosa-associated microbiota in the jejunum. Titanium dioxide was added in phase three diets as an indigestible marker. Ileal digesta was collected to measure apparent ileal digestibility (AID) of nutrients. Data were analyzed using MIXED and NLMIXED procedures of SAS. Increasing SPC supplementation by replacing animal protein supplements linearly decreased (P < 0.05) the BW, ADG, and ADFI of pigs during the overall period, and linearly increased (P < 0.05) peptide tyrosine tyrosine (PYY) in jejunum. Increasing SPC supplementation linearly decreased (P < 0.05) feed cost per weight gain. In the exponential model, SPC can replace animal protein supplements up to 10.5% and 16.5% without reducing the ADG and ADFI of pigs, respectively. The SPC 100% decreased (P < 0.05) Helicobacteraceae, Campylobacteraceae, alpha diversity, and changed beta diversity of microbiota in the jejunal mucosa. In conclusion, SPC supplementation replacing animal protein supplements reduced growth performance by reducing feed intake, which might be related to increased PYY. However, 10.5% and 16.8% of animal protein supplements can be replaced by SPC without affecting BW gain and feed intake of nursery pigs, respectively. Complete removal of animal protein supplements by SPC supplementation modulated the composition of jejunal mucosa-associated microbiota by reducing Helicobacteraceae and Campylobacteraceae, whereas without affecting the intestinal immune status, intestinal oxidative stress status, intestinal morphology, and AID of nutrients in nursery pigs.


Due to the high-quality nutrients and functional compounds, animal protein supplements are generally included in nursery pig diets to relieve the negative impacts caused by weaning stress. However, the high cost, short supply, and potential safety issues of animal protein supplements limit their use. Soybean meal is commonly used in swine diets due to the high nutritional values and competitive cost, however, antinutritional factors in soybean meal have been shown to impair the health and growth of nursery pigs. Soy protein concentrate is processed from soybean meal by ethanol extraction and efficiently removes the anti-nutritional factors. The aim of this study was to investigate the effects of soy protein concentrate replacing animal protein supplements at various levels on intestinal immune status, intestinal oxidative stress status, nutrient digestibility, and growth performance of nursery pigs. The use of soy protein concentrate completely replacing animal protein supplements showed benefits on modulating the bacterial ecosystem on the mucosal lining of the small intestine by decreasing potentially harmful bacteria, whereas without affecting intestinal immune status, intestinal oxidative stress status, intestinal morphology, and nutrient digestibility. However, excessive use of soy protein concentrate replacing animal protein supplements decreased the weight gain of nursery pigs due to reduced feed intake.


Asunto(s)
Microbiota , Proteínas de Soja , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Dieta/veterinaria , Suplementos Dietéticos/análisis , Digestión , Mucosa Intestinal/metabolismo , Nutrientes/metabolismo , Estrés Oxidativo , Péptidos/metabolismo , Proteínas de Soja/metabolismo , Porcinos , Tirosina/metabolismo , Aumento de Peso
7.
J Therm Biol ; 94: 102751, 2020 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-33292992

RESUMEN

The medicinal plant Artemisia annua L. is well known for its antimalarial compound artemisinin and the antioxidant capacity of its active ingredients. However, low bioavailability of Artemisia annua L. limits its therapeutic potential, fermentation of Artemisia annua L. can improve its bioavailability. This study was aimed to investigate the effects of dietary supplementation of enzymatically-treated Artemisia annua L. (EA) on reproductive performance, antioxidant status, milk composition of heat-stressed sows and intestinal barrier integrity of their preweaning offspring. 135 multiparous sows of average parity 4.65 (Landrace × large white) at day 85 of pregnancy were randomly distributed into 3 treatments. Sows in the control group were housed at control rooms (temperature: 27.12 ± 0.18 °C, temperature-humidity index (THI): 70.90 ± 0.80) and fed the basal diet. Sows in the HS, HS + EA groups were fed the basal diet supplemented with 0 or 1.0 g/kg EA respectively, and reared at heat stress rooms (temperature: 30.11 ± 0.16 °C, THI: 72.70 ± 0.60). Heat stress increased the malondialdehyde (MDA) content, reduced the activities of total antioxidant capacity (T-AOC) and total superoxide dismutase (T-SOD) of sows and piglets, and seriously compromised the antioxidant capacity of the sows and the intestinal integrity of their offspring. However, dietary supplementation of 1.0 g/kg EA reduced the MDA content, increased the activities of T-SOD and T-AOC in serum, colostrum, and milk of heat-stressed sows, and increased colostrum yield and 14-d milk fat content. EA supplementation also increased piglet weaning weight and the activities of T-SOD and T-AOC in serum. In addition, the abundances of intestinal tight junction proteins claudin-1 and occludin were up-regulated in piglets in EA-supplemented group. In conclusion, dietary EA supplementation at 1.0 g/kg can alleviate the oxidative stress in heat-stressed sows, improve the antioxidant capacity in both sows and their offspring, and promote the intestinal barrier integrity in their offspring. EA may be a potent dietary supplement that ameliorates oxidative stress in livestock production by improving the antioxidant capacity.


Asunto(s)
Artemisia annua , Suplementos Dietéticos , Calor/efectos adversos , Estrés Oxidativo , Reproducción , Alimentación Animal , Animales , Artemisia annua/química , Celulasa/química , Dieta/veterinaria , Femenino , Glutatión/sangre , Trastornos de Estrés por Calor/sangre , Trastornos de Estrés por Calor/genética , Trastornos de Estrés por Calor/veterinaria , Leche/química , Oxidorreductasas/sangre , Poligalacturonasa/química , Embarazo , Porcinos/sangre , Porcinos/genética , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/genética , Proteínas de Uniones Estrechas/genética
8.
Enzyme Microb Technol ; 126: 41-49, 2019 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-31000163

RESUMEN

The limited thermostability of Yarrowia lipolytica lipase 2 (Lip2) hampers its industrial application. To improve its thermostability, we combined single disulfide bonds which our group identified previously. In this study, combining different regional disulfide bonds had greater effect than combining same regional disulfide bonds. Furthermore, mutants with 4, 5, and 6 disulfide bonds exhibited dramatically enhanced thermostability. Compared with the wild-type, sextuple mutant 6s displayed a 22.53 and 31.23 ℃ increase in the melting temperature (Tm) and the half loss temperature at 15 min (T15 50), respectively, with greater pH stability and a wider reaction pH range. Molecular dynamics simulation revealed that multiple disulfide bonds resulted in more rigid structures of mutants 4s, 5s and 6s, and prolonged enzyme unfolding times. Moreover, secretions of mutants 5s and 6s were significantly increased by 60% and 80% by co-expressing with the chaperone protein disulfide isomerase (PDI), which mitigated the reduced production issue caused by multiple disulfide bonds. Results of this study indicated that enhanced heat endurance giving more potential for industrial application.


Asunto(s)
Hidrolasas de Éster Carboxílico/química , Hidrolasas de Éster Carboxílico/metabolismo , Disulfuros/química , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Temperatura , Yarrowia/enzimología , Hidrolasas de Éster Carboxílico/genética , Estabilidad de Enzimas , Proteínas Fúngicas/genética , Simulación de Dinámica Molecular , Mutagénesis Sitio-Dirigida , Mutación , Conformación Proteica , Proteína Disulfuro Isomerasas/metabolismo , Factores de Transcripción/metabolismo
9.
Artículo en Inglés | MEDLINE | ID: mdl-29644075

RESUMEN

BACKGROUND: Milk protein is crucial for milk quality in sows and health of newborn piglets. Plasma amino acids (AA) in sows are important precursors for milk protein synthesis in the mammary gland. In order to study the regulation of AA transported in sow mammary glands and possible underlying mechanisms, we measured the expression of genes coding for milk proteins, AA transporter expressions, and plasma AA concentrations in sows at three different physiological stages (D-17, D1 and D17 of lactation), and then further investigated the regulation of AA transport across the cell membrane by adaptive mechanisms using pig mammary epithelial cells (PMEC) as an in vitro model. PMEC were cultured in DMEM:F12 with 4 amino acid concentrations (0 × AA complex, 1 × AA complex, 5 × AA complex, and 25 × AA complex). Classes of AA complexes evaluated in this study included neutral AAs (L-Ala + L-Ser + L-Cys), acidic AAs (L-Asp, L-Glu) and neutral + basic AAs (L-Ala + L-Ser + L-Cys + L-Lys). RESULTS: Our results indicated that mRNA expression of genes coding for milk protein (αs1-casein, αs2-casein, ß-casein and κ-casein) increased significantly with the advance of physiological stage (P < 0.05), and plasma concentrations of most AAs including threonine, serine, glutamate, alanine, valine, cysteine, methionine, isoleucine and tyrosine were greater at D1 of lactation compared with D-17 and D17 of lactation (P < 0.05). Additionally, protein and gene expressions of AA transporters including excitatory AA transporter 3 (EAAT3), alanine/serine/cysteine/threonine transporter (ASCT1) and sodium-coupled neutral AA transporter 1 (SNAT2) were greater in lactating sow mammary glands compared with sow mammary glands in late pregnancy (P < 0.05). The mRNA expressions of SLC38A2, SLC1A1, SLC6A14 increased significantly in the cell mediums supplemented with 5 × and 25 × of AA complexes compared with those cells cultured in DMEM/F12 cell medium (P < 0.05). The mRNA expressions of SLC38A, SLC1A4, and SLC6A14 also increased in EBSS cell medium compared to DMEM/F12. However, only mRNA expression of SLC38A decreased when AA complex was added into EBSS (P < 0.05). CONCLUSION: AA transportation was positively regulated in sow mammary glands with the advance of physiological stage from late pregnancy to peak of lactation and AA transporters in PMECs were adaptively regulated by changed AA concentrations.

10.
Trop Anim Health Prod ; 50(5): 973-982, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29504061

RESUMEN

A 2 × 2 factorial arrangement (rearing room with or without pad-fan cooling × diet with or without 2.5 kg/t organic acid) was used to evaluate the effect of pad-fan cooling and dietary organic acid supplementation during perinatal period on reproductive performance and antioxidant status of sows in hot weather. This study was conducted in a subtropical city in Guangdong Province in South China between August and October, 2015. At day 85 of gestation, a total of 112 sows were randomly assigned to the four treatments with 28 sows per treatment, and maintained until day 21 of lactation, and the feeding trial lasted for 51 days. During the experimental period, room temperature and humidity were recorded hourly. The lactation feed intake of sows (P = 0.109) and stillbirths (P < 0.05) increased when the sows were reared in the room with the pad-fan cooling against the room without pad-fan cooling. The number of weak newborns per litter and the malondialdehyde content in days 14 and 21 milk decreased (P < 0.05), while the lactation feed intake of sows, weaned litter weights, and individual pig weights increased when the sows were fed the organic acid (P < 0.05). In conclusion, pad-fan cooling in rearing room improved the lactation feed intake of sows, and dietary organic acid supplementation improved reproductive performance and milk antioxidant status of sows. Pad-fan cooling is recommended in farrowing room, but not in gestating room.


Asunto(s)
Alimentación Animal/análisis , Antioxidantes/química , Dieta/veterinaria , Calor , Lactancia/efectos de los fármacos , Reproducción/efectos de los fármacos , Ácidos/uso terapéutico , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , China , Suplementos Dietéticos , Femenino , Malondialdehído/química , Leche , Paridad , Embarazo , Porcinos , Destete , Tiempo (Meteorología)
11.
Arch Anim Nutr ; 71(6): 441-454, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-29110578

RESUMEN

Two experiments were conducted to investigate the effect of combined supplementation of α-galactosidase and xylanase on nutrient digestibility and growth performance in growing pigs. Experiment 1 had a 2 × 2 Latin square design, where eight barrows (45.0 ± 0.52 kg body weight [BW]) were fitted with a simple T-cannula in the distal ileum and received a basal diet without or with supplementation of α-galactosidase (12 U/kg diet) and xylanase (15 AXC/kg diet) within two periods of 10 d. The apparent ileal digestibility (AID) and apparent total tract digestibility of nutrients, pH, viscosity of digesta and digestive enzyme activities were assessed. In Experiment 2, a total of 432 growing pigs (initial BW 44.7 ± 0.66 kg) were allocated to four treatments. Diets were based on corn and soybean meal and had a normal or reduced nutrient level (reduced by 0.42 kJ digestible energy [DE] per kg and 0.8% crude protein). Both diets were offered without or with supplementation of α-galactosidase and xylanase. The growth performance was assessed within a 43-d feeding period, where at the end, biochemical serum indices were estimated. In Experiment 1, the enzyme-supplemented diet had a greater contents of DE and DE/gross energy ratio (p < 0.05), and a higher AID of Arg, raffinose, stachyose and arabinoxylan (p < 0.05). In Experiment 2, the low nutrient level caused lower daily gain (p < 0.05), which was partially compensated by enzyme addition. Enzyme addition also increased the serum concentration of Lys (p < 0.05). Moreover, it appears that the tested enzyme supplementation could increase dietary DE, serum total amino acid concentrations and decrease serum urea nitrogen.


Asunto(s)
Dieta/veterinaria , Digestión/efectos de los fármacos , Endo-1,4-beta Xilanasas/metabolismo , Sus scrofa/fisiología , alfa-Galactosidasa/metabolismo , Alimentación Animal/análisis , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Suplementos Dietéticos/análisis , Endo-1,4-beta Xilanasas/administración & dosificación , Femenino , Masculino , Glycine max/química , Sus scrofa/crecimiento & desarrollo , Zea mays/química , alfa-Galactosidasa/administración & dosificación
12.
Protein Expr Purif ; 138: 34-45, 2017 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-28583876

RESUMEN

Relatively poor heterologous protein yields have limited the commerical applications of Galactomyces geotrichum lipase I (GGl I) efficacy trials. To address this, we have redesigned the GGl I gene to preferentially match codon frequencies of Pichia pastoris (P. pastoris) while retaining the same amino acid sequence. The wild type and codon optimised GGl I (GGl I-wt and GGl I-op) were synthesised and cloned into pPICZαA with an N-terminal 6 × His tag sequence and expressed in P. pastoris X 33. The hydrolytic activity of GGl I-op was 150 U/mL, whereas the activity of the GGl I-wt could not be detected. GGl I-op recombinant proteins were purified by Ni-affinity chromatography and then characterised. The identity and purity of GGl I were confirmed by SDS-PAGE, MALDI-TOF mass spectrometry and Western blot analysis. Enzymatic deglycosylation was used to show that the lipase is a glycosylated protein, containing ∼10% sugar. The molecular weight (MW) of the GGl I secreted by recombinant P. pastoris was approximated at 63 kDa. The optimum pH and temperature of the recombinant lipase were 8.0 and 35 °C, respectively. The enzyme was active over a broad pH range (7.0-9.0) and temperature range (20 °C-45 °C). The lipase showed high activity toward medium- and long-chain fatty acid methyl esters (C8-C16) and retained much of its activity in the presence of Tween-80 and Trition X-100. Lipase activity was stimulated by Mg2+, Ca2+, Mn2+ and Cu2+ and inhibited by Fe2+, Fe3+, Zn2+ and Co2+. This lipase may prove useful to the detergent industry and in organic synthesis reactions.


Asunto(s)
Codón/metabolismo , Ácidos Grasos/metabolismo , Proteínas Fúngicas/metabolismo , Lipasa/metabolismo , Pichia/genética , Proteínas Recombinantes de Fusión/metabolismo , Saccharomycetales/química , Secuencia de Aminoácidos , Cationes Bivalentes , Cromatografía de Afinidad , Clonación Molecular , Codón/química , Pruebas de Enzimas , Ésteres , Proteínas Fúngicas/genética , Proteínas Fúngicas/aislamiento & purificación , Expresión Génica , Vectores Genéticos/química , Vectores Genéticos/metabolismo , Histidina/genética , Histidina/aislamiento & purificación , Histidina/metabolismo , Concentración de Iones de Hidrógeno , Cinética , Lipasa/genética , Lipasa/aislamiento & purificación , Metales/metabolismo , Peso Molecular , Oligopéptidos/genética , Oligopéptidos/aislamiento & purificación , Oligopéptidos/metabolismo , Pichia/metabolismo , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificación , Saccharomycetales/enzimología , Alineación de Secuencia , Especificidad por Sustrato , Temperatura
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