On the ensiling of lucerne by means of lactic acid fermentation

Orla-Jensen, S.; Orla-Jensen, Anna D.; Kjaer, Agnete

doi:10.1007/bf02272656

Cited by 12 publications

(7 citation statements)

References 2 publications

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“…Based on the annotation of L. plantarum WCFS1 we found one mrr gene (GeneID:1061548). This gene (99% sequence identity) was also present in the genome sequences of L. plantarum JDM1 and of the type strain L. plantarum DSM20174 [39]; no putative mrr genes were detected in the genome of L. plantarum ST-III. BLAST analysis [40] of these genes revealed further identities (60% on amino acid level) to Lactobacillus farciminis KCTC 3681 and Lactobacillus reuteri 100-23, all other identities were below 60%.…”

Section: Resultsmentioning

confidence: 99%

“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”

2012

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BackgroundLactic acid bacteria (LAB) play an important role in agricultural as well as industrial biotechnology. Development of improved LAB strains using e.g. library approaches is often limited by low transformation efficiencies wherefore one reason could be differences in the DNA methylation patterns between the Escherichia coli intermediate host for plasmid amplification and the final LAB host. In the present study, we examined the influence of DNA methylation on transformation efficiency in LAB and developed a direct cloning approach for Lactobacillus plantarum CD033. Therefore, we propagated plasmid pCD256 in E. coli strains with different dam/dcm-methylation properties. The obtained plasmid DNA was purified and transformed into three different L. plantarum strains and a selection of other LAB species.ResultsBest transformation efficiencies were obtained using the strain L. plantarum CD033 and non-methylated plasmid DNA. Thereby we achieved transformation efficiencies of ~ 109 colony forming units/μg DNA in L. plantarum CD033 which is in the range of transformation efficiencies reached with E. coli. Based on these results, we directly transformed recombinant expression vectors received from PCR/ligation reactions into L. plantarum CD033, omitting plasmid amplification in E. coli. Also this approach was successful and yielded a sufficient number of recombinant clones.ConclusionsTransformation efficiency of L. plantarum CD033 was drastically increased when non-methylated plasmid DNA was used, providing the possibility to generate expression libraries in this organism. A direct cloning approach, whereby ligated PCR-products where successfully transformed directly into L. plantarum CD033, obviates the construction of shuttle vectors containing E. coli-specific sequences, as e.g. a ColEI origin of replication, and makes amplification of these vectors in E. coli obsolete. Thus, plasmid constructs become much smaller and occasional structural instability or mutagenesis during E. coli propagation is excluded. The results of our study provide new genetic tools for L. plantarum which will allow fast, forward and systems based genetic engineering of this species.

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Section: Resultsmentioning

confidence: 99%

“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”

2012

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“…Since there is evidence of qualitative changes in the microflora of silage as fermentation proceeds (Orla-Jensen et at., 1947;Kempton and San Clemente, 1.959, Langston et al, 1962;Moon, 1981 and Moon et at,, 1981a) a mixed inoculum of streptococci and other genera of lactic acid bacteria is more likely to influence the outcome than a single culture, "i he results relating to the streptococci, lactobacilli and pediococci were therefore considered in this context. Moreover, such an approach greatly assisted in the interpretation of the large volume of data generated.…”

Section: Discussioqmentioning

confidence: 99%

“…This point, together with the knowledge that within the silage microllora inelficient acid-producing bacteria may be in a majority (Langston^r at,, 1962), highlights the need for the application of sufficient viable organisms lo ensure their domination over the indigenous microflora. There is evidence to show that laclic acid bacteria belonging to the genera Streptococcus and Leuconostoc initiate fermentation and are superseded by species of Lactobaciltus and Pediococcus (Orla-Jensen et al, 1947;Kempton and San Clemenle, 1959;Langston et at,. 1962;Moon, 1981 andMoon et at.,, 1981a).…”

Section: Introductionmentioning

confidence: 99%

An investigation into the effect of cultures of lactic acid bacteria on fermentation in silage

Woolford¹,

Sawczyc

1984

Grass and Forage Science

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Twenty-one strains of lactic acid bacteria were subjected to a range of tests based on established criteria lo ascertain their suitability for use as inoculants for silage. The tests included a monitor of their growth in the presence of representatives of the microflora of forage crops with which they might have to compete for substrate, and their ability to effect rapid acidification and achieve a low pH in conditions which simulate low and high ensiling potential. In addition, their ability to produce acid from sugars and their action against organic acids found in forage crops, together with their tolerance of acid, elevated temperatures and conditions of low moisture availability (e.g. in wilted material) and possible proteolytic and amylolytic activities were examined. None of these cultures satisfied all of these criteria. However, three strains {Streptococcus durans str. 1024, Lactobacillus acidophilus, str. 2356 and Lactohacitlus plantarum str. 6) had a greater potential than the others.

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“…Belonging to class Actinobacteria, order Actinomycetales and family Microbacteriaceae, the genus Microbacterium was first proposed by Orla-Jensen with Microbacterium lacticum as the type species [1]. At the time of writing, the genus Microbacterium comprised 139 recognized species with validly published names (LPSN: www.bacterio.net).…”

Section: Introductionmentioning

confidence: 99%

Microbacterium abyssi sp. nov. and Microbacterium limosum sp. nov., two new species of the genus Microbacterium, isolated from deep-sea sediment samples

Ding,

Chen

et al. 2024

International Journal of Systematic and Evolutionary Microbiology

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Two Gram-positive, non-motile, short rod-shaped actinomycete strains, designated as A18JL241T and Y20T, were isolated from deep-sea sediment samples collected from the Southwest Indian Ocean and Western Pacific Ocean, respectively. Both of the isolates were able to grow within the temperature range of 5–40 °C, NaCl concentration range of 0–7 % (w/v) and at pH 6.0–12.0. The two most abundant cellular fatty acids of both strains were anteiso-C15 : 0 and anteiso-C17 : 0. The major polar lipid contents of the two strains were phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and one unidentified glycolipid. These two strains shared common chemotaxonomic features comprising MK-10 and MK-12 as the respiratory quinones. The genomic DNA G+C contents of the two strains were 68.1 and 70.4 mol%, respectively. The 16S rRNA gene phylogeny showed that the novel strains formed two distinct sublines within the genus Microbacterium. Strain A18JL241T was most closely related to the type strain of Microbacterium tenebrionis KCTC 49593T (98.8 % sequence similarity), whereas strain Y20T formed a tight cluster with the type strain of Microbacterium schleiferi NBRC 15075T (99.0 %). The orthologous average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values with the type strains of related Microbacterium species were in the range of 74.1–89.1 % and 19.4–36.9 %, respectively, which were below the recognized thresholds of 95–96 % ANI and 70 % dDDH for species definition. Based on the results obtained here, it can be concluded that strains A18JL241T and Y20T represent two novel species of the genus Microbacterium, for which the names Microbacterium abyssi sp. nov. (type strain A18JL241T=JCM 33956T=MCCC 1A16622T) and Microbacterium limosum sp. nov. (type strain Y20T=JCM 33960T=MCCC 1A16747T) are proposed.

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On the ensiling of lucerne by means of lactic acid fermentation

Cited by 12 publications

References 2 publications

“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”

“Direct cloning in Lactobacillus plantarum: Electroporation with non-methylated plasmid DNA enhances transformation efficiency and makes shuttle vectors obsolete”

An investigation into the effect of cultures of lactic acid bacteria on fermentation in silage

Microbacterium abyssi sp. nov. and Microbacterium limosum sp. nov., two new species of the genus Microbacterium, isolated from deep-sea sediment samples

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