scholarly journals Impaired Growth Rates in Milk of Lactobacillus helveticus Peptidase Mutants Can Be Overcome by Use of Amino Acid Supplements

2003 ◽  
Vol 185 (11) ◽  
pp. 3297-3306 ◽  
Author(s):  
Jeffrey E. Christensen ◽  
James L. Steele

ABSTRACT To evaluate the contribution of intracellular peptidases to the growth of the 14-amino-acid (aa) auxotroph Lactobacillus helveticus CNRZ32, single- and multiple-peptidase-deletion mutants were constructed. Two broad-specificity aminopeptidases (PepC and PepN) and X-prolyl dipeptidyl aminopeptidase (PepX) were inactivated through successive cycles of chromosomal gene replacement mutagenesis. The inactivation of all three peptidases in JLS247 (ΔpepC ΔpepN ΔpepX) did not affect the growth rate in amino acid-defined medium. However, the peptidase mutants generally had decreased specific growth rates when acquisition of amino acids required hydrolysis of the proteins in milk, the most significant result being a 73% increase in generation time for JLS247. The growth rate deficiencies in milk were overcome by amino acid supplements with some specificity to each of the peptidase mutants. For example, milk supplementation with Pro resulted in the most significant growth rate increase for ΔpepX strains and a 7-aa supplement (Asn, Cys, Ile, Pro, Ser, Thr, and Val) resulted in a JLS247 growth rate indistinguishable from that of the wild type. Our results show that characterization of the activities of the broad-specificity aminopeptidases had little predictive value regarding the amino acid supplements found to enhance the milk growth rates of the peptidase mutant strains. These results represent the first determination of the physiological roles with respect to specific amino acid requirements for peptidase mutants grown in milk.

2001 ◽  
Vol 67 (4) ◽  
pp. 1846-1850 ◽  
Author(s):  
Elvira M. Hebert ◽  
Graciela S. De Giori ◽  
Raul R. Raya

ABSTRACT A slowly milk-coagulating variant (Fmc−) ofLactobacillus helveticus CRL 1062, designated S1, was isolated and characterized. Strain S1 possessed all the known essential components required to utilize casein as a nitrogen source, which include functional proteinase and peptidase activities as well as functional amino acid, di- and tripeptide, and oligopeptide transport systems. The amino acid requirements of strain S1 were similar to those of the parental strain. However, on a purine-free, chemically defined medium, the growth rate of the Fmc− strain was threefold lower than that of the wild-type strain. L. helveticusS1 was found to be defective in IMP dehydrogenase activity and therefore was deficient in the ability to synthesize XMP and GMP. This conclusion was further supported by the observation that the addition of guanine or xanthine to milk, a substrate poor in purine compounds, restored the Fmc+ phenotype of L. helveticusS1.


2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Kanhaiya Kumar ◽  
Vishwesh Venkatraman ◽  
Per Bruheim

Abstract Background Saccharomyces cerevisiae is a well-known popular model system for basic biological studies and serves as a host organism for the heterologous production of commercially interesting small molecules and proteins. The central metabolism is at the core to provide building blocks and energy to support growth and survival in normal situations as well as during exogenous stresses and forced heterologous protein production. Here, we present a comprehensive study of intracellular central metabolite pool profiling when growing S. cerevisiae on different carbon sources in batch cultivations and at different growth rates in nutrient-limited glucose chemostats. The latest versions of absolute quantitative mass spectrometry-based metabolite profiling methodology were applied to cover glycolytic and pentose phosphate pathway metabolites, tricarboxylic acid cycle (TCA), complete amino acid, and deoxy-/nucleoside phosphate pools. Results Glutamate, glutamine, alanine, and citrate were the four most abundant metabolites for most conditions tested. The amino acid is the dominant metabolite class even though a marked relative reduction compared to the other metabolite classes was observed for nitrogen and phosphate limited chemostats. Interestingly, glycolytic and pentose phosphate pathway (PPP) metabolites display the largest variation among the cultivation conditions while the nucleoside phosphate pools are more stable and vary within a closer concentration window. The overall trends for glucose and nitrogen-limited chemostats were increased metabolite pools with the increasing growth rate. Next, comparing the chosen chemostat reference growth rate (0.12 h−1, approximate one-fourth of maximal unlimited growth rate) illuminates an interesting pattern: almost all pools are lower in nitrogen and phosphate limited conditions compared to glucose limitation, except for the TCA metabolites citrate, isocitrate and α-ketoglutarate. Conclusions This study provides new knowledge-how the central metabolism is adapting to various cultivations conditions and growth rates which is essential for expanding our understanding of cellular metabolism and the development of improved phenotypes in metabolic engineering.


PeerJ ◽  
2015 ◽  
Vol 3 ◽  
pp. e1392
Author(s):  
Juan C. Levesque

Ladyfish (Elopssp) are a common and economically valuable coastal nearshore species found along coastal beaches, bays, and estuaries of the southeastern United States, and subtropical and tropical regions worldwide. Previously, ladyfish were a substantial bycatch in Florida’s commercial fisheries, but changes in regulations significantly reduced commercial landings. Today, ladyfish are still taken in commercial fisheries in Florida, but many are also taken by recreational anglers. Life-history information and research interest in ladyfish is almost non-existent, especially information on age and growth. Thus, the overarching purpose of this study was to expand our understanding of ladyfish age and growth characteristics. The specific objectives were to describe, for the first time, age, growth, and recruitment patterns of juvenile ladyfish from the east coast of Florida (USA). In the Indian River Lagoon (IRL), annual monthly length-frequency distributions were confounded because a few small individuals recruited throughout the year; monthly length-frequency data generally demonstrated a cyclical pattern. The smallest were collected in September and the largest in May. Post-hoc analysis showed no significant difference in length between August and May, or among the other months. In Volusia County (VC), annual monthly length-frequency distribution demonstrated growth generally occurred from late-winter and spring to summer. The smallest ladyfish were collected in February and the largest in August. On average, the absolute growth rate in the IRL was 36.3 mm in 60 days or 0.605 mm day−1. Cohort-specific daily growth rates, elevations, and coincidentals were similar among sampling years. Cohort-specific growth rates ranged from 1.807 in 1993 to 1.811 mm day−1in 1994. Overall, growth was best (i.e., goodness of fit) described by exponential regression. On average, the absolute growth rate in VC was 28 mm in 150 days or 0.1866 mm day−1. Cohort-specific daily growth rates were significantly different among sampling years; however, the elevations and coincidentals were similar. Cohort-specific growth rates ranged from 1.741 in 1994 to 1.933 mm day−1in 1993. Mean ladyfish growth was best described by linear regression; however, natural growth was explained better by exponential regression. In the IRL, the corrected exponential growth equation yielded a size-at-age 1 of 156.0 mm SL, which corresponded to an estimated growth rate of 0.4356 mm day−1. In VC, the corrected exponential growth equation yielded a size-at-age 1 of 80 mm SL corresponding to an estimated growth rate of 0.2361 mm day−1.


2013 ◽  
Vol 10 (8) ◽  
pp. 5267-5280 ◽  
Author(s):  
F. H. Chang ◽  
E. C. Marquis ◽  
C. W. Chang ◽  
G. C. Gong ◽  
C. H. Hsieh

Abstract. Allometric scaling of body size versus growth rate and mortality has been suggested to be a universal macroecological pattern, as described by the metabolic theory of ecology (MTE). However, whether such scaling generally holds in natural assemblages remains debated. Here, we test the hypothesis that the size-specific growth rate and grazing mortality scale with the body size with an exponent of −1/4 after temperature correction, as MTE predicts. To do so, we couple a dilution experiment with the FlowCAM imaging system to obtain size-specific growth rates and grazing mortality of natural microphytoplankton assemblages in the East China Sea. This novel approach allows us to achieve highly resolved size-specific measurements that would be very difficult to obtain in traditional size-fractionated measurements using filters. Our results do not support the MTE prediction. On average, the size-specific growth rates and grazing mortality scale almost isometrically with body size (with scaling exponent ∼0.1). However, this finding contains high uncertainty, as the size-scaling exponent varies substantially among assemblages. The fact that size-scaling exponent varies among assemblages prompts us to further investigate how the variation of size-specific growth rate and grazing mortality can interact to determine the microphytoplankton size structure, described by normalized biomass size spectrum (NBSS), among assemblages. We test whether the variation of microphytoplankton NBSS slopes is determined by (1) differential grazing mortality of small versus large individuals, (2) differential growth rate of small versus large individuals, or (3) combinations of these scenarios. Our results indicate that the ratio of the grazing mortality of the large size category to that of the small size category best explains the variation of NBSS slopes across environments, suggesting that higher grazing mortality of large microphytoplankton may release the small phytoplankton from grazing, which in turn leads to a steeper NBSS slope. This study contributes to understanding the relative importance of bottom-up versus top-down control in shaping microphytoplankton size structure.


1998 ◽  
Vol 64 (11) ◽  
pp. 4226-4233 ◽  
Author(s):  
Pim Van Hoek ◽  
Johannes P. Van Dijken ◽  
Jack T. Pronk

ABSTRACT The specific growth rate is a key control parameter in the industrial production of baker’s yeast. Nevertheless, quantitative data describing its effect on fermentative capacity are not available from the literature. In this study, the effect of the specific growth rate on the physiology and fermentative capacity of an industrialSaccharomyces cerevisiae strain in aerobic, glucose-limited chemostat cultures was investigated. At specific growth rates (dilution rates, D) below 0.28 h−1, glucose metabolism was fully respiratory. Above this dilution rate, respirofermentative metabolism set in, with ethanol production rates of up to 14 mmol of ethanol · g of biomass−1 · h−1at D = 0.40 h−1. A substantial fermentative capacity (assayed offline as ethanol production rate under anaerobic conditions) was found in cultures in which no ethanol was detectable (D < 0.28 h−1). This fermentative capacity increased with increasing dilution rates, from 10.0 mmol of ethanol · g of dry yeast biomass−1 · h−1 at D= 0.025 h−1 to 20.5 mmol of ethanol · g of dry yeast biomass−1 · h−1 atD = 0.28 h−1. At even higher dilution rates, the fermentative capacity showed only a small further increase, up to 22.0 mmol of ethanol · g of dry yeast biomass−1 · h−1 at D= 0.40 h−1. The activities of all glycolytic enzymes, pyruvate decarboxylase, and alcohol dehydrogenase were determined in cell extracts. Only the in vitro activities of pyruvate decarboxylase and phosphofructokinase showed a clear positive correlation with fermentative capacity. These enzymes are interesting targets for overexpression in attempts to improve the fermentative capacity of aerobic cultures grown at low specific growth rates.


2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 75-75
Author(s):  
Laura L Greiner

Abstract Over the last 25 years, the sow has developed to have a leaner body mass to address the need for leaner, fast growing offspring. The leaner body mass results in a female that biologically can have a lower feed intake. Furthermore, the number of pigs born per litter and milk production have also increased during the same timeframe. Much research has been conducted over the years to investigate the amino acid and energy requirements of both the lactating and gestating sow; however, application of the research has resulted in varied responses. The varied responses can be related to sow health, environmental influences, litter growth rates, and anticipated body reserve changes during lactation. Furthermore, calculations associated with the regression of reproductive tissue after farrowing can complicate the analysis of amino acid requirements. This presentation will discuss some of the recent research associated with feeding the modern sow, potentially challenging some of the differences in data, and offering some thoughts on looking towards the next 25 years of sow production.


1950 ◽  
Vol 28c (1) ◽  
pp. 1-6 ◽  
Author(s):  
R. H. Wallace ◽  
A. G. Lochhead

A study was made of the more specific amino acid requirements of bacteria from the rhizospheres of clover, flax, and wheat plants for which a chemically defined medium containing 23 amino acids provided essentials for maximum growth. Of seven groups of amino acids, the sulphur-containing group (cysteine, methionine, and taurine) was found to be of special significance, the omission of this group resulting in a pronounced decrease in the percentage of organisms able to develop. Further study of organisms dependent upon this group of amino acids for growth showed methionine to be by far the most essential compound. While evident for bacteria from the rhizosphere of all three crops, the effect was more pronounced in the case of clover than with flax or wheat.


Microbiology ◽  
2005 ◽  
Vol 151 (1) ◽  
pp. 135-143 ◽  
Author(s):  
Tiina M. Pakula ◽  
Katri Salonen ◽  
Jaana Uusitalo ◽  
Merja Penttilä

Trichoderma reesei was cultivated in chemostat cultures on lactose-containing medium. The cultures were characterized for growth, consumption of the carbon source and protein production. Secreted proteins were produced most efficiently at low specific growth rates, 0·022–0·033 h−1, the highest specific rate of total protein production being 4·1 mg g−1 h−1 at the specific growth rate 0·031 h−1. At low specific growth rates, up to 29 % of the proteins produced were extracellular, in comparison to only 6–8 % at high specific growth rates, 0·045–0·066 h−1. To analyse protein synthesis and secretion in more detail, metabolic labelling of proteins was applied to analyse production of the major secreted protein, cellobiohydrolase I (CBHI, Cel7A). Intracellular and extracellular labelled CBHI was quantified and analysed for pI isoforms in two-dimensional gels, and the synthesis and secretion rates of the molecule were determined. Both the specific rates of CBHI synthesis and secretion were highest at low specific growth rates, the optimum being at 0·031 h−1. However, at low specific growth rates the secretion rate/synthesis rate ratio was significantly lower than that at high specific growth rates, indicating that at low growth rates the capacity of cells to transport the protein becomes limiting. In accordance with the high level of protein production and limitation in the secretory capacity, the transcript levels of the unfolded protein response (UPR) target genes pdi1 and bip1 as well as the gene encoding the UPR transcription factor hac1 were induced.


1978 ◽  
Vol 24 (1) ◽  
pp. 28-30 ◽  
Author(s):  
Adrian P. Wills ◽  
E. C. S. Chan

When deprived of biotin, Arthrobacter globiformis 425 exhibits abnormal morphology (large, branched forms of variable size) and a retardation of its normal growth rate. In chemostat cultures, when cells were grown under glucose limitation, the morphology was normal (coccoids or rods) at specific growth rates between 0.05 and 0.125 h−1 (doubling times between 14 and 5.5 h, respectively) at 25 °C. The coccoid-to-rod morphogenesis occurs at a specific growth rate of 0.11 h−1. At the same specific growth rates and temperature, but under biotin limitation, abnormal morphology was observed.


Sign in / Sign up

Export Citation Format

Share Document