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. 2016 Aug 26;11(1):54. doi: 10.1186/s40793-016-0176-4

First high quality draft genome sequence of a plant growth promoting and cold active enzyme producing psychrotrophic Arthrobacter agilis strain L77

Ram N Singh 1, Sonam Gaba 1, Ajar N Yadav 1, Prakhar Gaur 1, Sneha Gulati 1, Rajeev Kaushik 1, Anil K Saxena 1,2,✉
PMCID: PMC5000428  PMID: 27570579

Abstract

Arthrobacter agilis strain L77, is a plant growth promoting and cold active hydrolytic enzymes producing psychrotrophic bacterium, isolated from Pangong Lake, a subglacial lake in north western Himalayas, India. Genome analysis revealed metabolic versatility with genes involved in metabolism and cold shock adaptation, utilization and biosynthesis of diverse structural and storage polysaccharides such as plant based carbon polymers. The genome of Arthrobacter agilis strain L77 consists of 3,608,439 bp (3.60 Mb) of a circular chromosome. The genome comprises of 3316 protein coding genes and 74 RNA genes, 725 hypothetical proteins, 25 pseudo-genes and 1404 unique genes.

Electronic supplementary material

The online version of this article (doi:10.1186/s40793-016-0176-4) contains supplementary material, which is available to authorized users.

Keywords: Arthrobacter, Psychrotrophic, PGPB, Cold-active enzymes, Pangong Lake, Himalayas

Introduction

The microorganisms from extreme environments are of particular importance in global ecology since the majority of terrestrial and aquatic ecosystems of our planet are permanently or seasonally submitted to cold temperatures. Microorganisms capable of coping with low temperatures are widespread in these natural environments where they often represent the dominant flora and they should therefore be regarded as the most successful colonizers of our planet. Members of the genus Arthrobacter [1, 2] are Gram-positive, show rods in exponential growth and cocci in their stationary phase, able to grow under aerobic as well as anaerobic conditions and belong to the phylum Actinobacteria [3]. Different species of Arthrobacter [1, 2] have been implicated in plant growth promotion [4], production of industrially important enzymes [5, 6] and as xeroprotectant [7, 8]. These reports suggest that species from Arthrobacter [1, 2] harbor genes for coding enzymes that can be useful in the industry, agriculture and biotechnology. Arthrobacter agilis [9] strain L77 was isolated from Pangong Lake, a subglacial lake in north western Himalayas, India and exhibit plant growth promoting attributes as well as production of hydrolytic enzymes. The culture was further characterized for production of EPS and anti-freeze compounds (AFCs). Here, we present the draft genome sequence of Arthrobacter agilis [9] strain L77 along with the description of genome properties and annotation.

Organism information

Classification and features

Arthrobacter agilis [9] strain L77 was isolated from frozen sub-glacial Pangong Lake (33°82′55.59″N and 78°59′26.69″E) in north western Himalaya, India (Table 1). This psychrotrophic bacterium was isolated using standard serial dilution method on Trypticase soya agar [10] plate and has been reported to possess plant growth promoting attributes and could produce cold active enzymes and AFCs. It could solubilize phosphorus, zinc and could produce indole acetic acid and ammonia. It could produce cold active enzymes such as lipase, amylase, protease, chitinase and β-galactosidase.

Table 1.

Classification and general features of Arthrobacter agilis strain L77

MIGS ID Property Term Evidence codea
Classification Domain Bacteria TAS [12]
Phylum Actinobacteria TAS [3]
Class Actinobacteria TAS [13]
Order Actinomycetales TAS [2, 14]
Family Micrococcaceae TAS [2, 15]
Genus Arthrobacter TAS [1, 2]
Species Arthrobacter agilis TAS [9]
Strain L77 NAS
Gram stain Positive IDA
Cell shape Polymorphic: Coccus to rod shaped IDA
Motility Non-motile TAS [9]
Sporulation Non-sporulating TAS [9]
Temperature range −10 °C −30 °C IDA
Optimum temperature 15 °C IDA
pH range; Optimum 6–9, 7 IDA
Carbon source Yeast extract, glucose, lactose, mannose TAS [9]
MIGS-6 Habitat Sub-glacial Lake IDA
MIGS-6.3 Salinity Grown on 5 % > NaCl (w/v) IDA
MIGS-22 Oxygen requirement Aerobic TAS [9]
MIGS-15 Biotic relationship Free living TAS [9]
MIGS-14 Pathogenicity Non-pathogeneic NAS
MIGS-4 Geographic location India, Leh Ladakh, Jammu & Kashmir TAS [10]
MIGS-5 Sample collection March 28, 2010 IDA
MIGS-4.1 Latitude 33°82′55.59″N NAS
MIGS-4.2 Longitude 78°59′26.69″E NAS
MIGS-4.4 Altitude 3215 m NAS
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aEvidence codes - TAS Traceable Author Statement (i.e., a direct report exists in the literature), NAS Non-traceable Author Statement (i.e., not directly observed for the living, isolated sample, but based on a generally accepted property for the species, or anecdotal evidence). These evidence codes are from the Gene Ontology project [49]

Strain L77 is a bright yellow colored (Fig. 1) Gram-positive, aerobic, non-motile bacterium exhibiting a rod-coccus cycle. The initial validation of bacterium was done by 16S rRNA gene sequencing using the universal eubacterial primers pA (5′-AGAGTTTGATCCTGGCTCAG-3′) and pH (5′-AAGGAGGTGATCCAGCCGCA-3′) [11]. The 16S rRNA gene sequence places Arthrobacter agilis strain L77 in the domain Bacteria [12] (Table 1), phylum Actinobacteria [3] and Class Actinobacteria [13], order Actinomycetales [2, 14] and family Micrococcaceae [2, 15] during homology search by BLAST [16]. Only few of the closely related species after reclassification [17] of genus Arthrobacter [1, 2,] with validly published names: A. agilisDSM 20550T [9], A. woluwensis 1551TDSM 10495 [18], A. methylotrophusDSM 14008T [19], A. tectiLMG 22282T [20], A. parietisLMG 22281T [20], A. subterraneus CH7TDSM 17585 [21], A. tumbaeLMG 19501T [20], Arthrobacter oryzae KV-651TDSM 25586 [22], Arthrobacter alkaliphilus LC6TDSM 23368 [23], Arthrobacter flavusJCM 11496T [24], A. cupressi D48TDSM 24664 [25], A. globiformisDSM 20124T [1, 2] were selected for drawing the phylogenetic position of strain L77.

Fig. 1.

Fig. 1

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Full grown yellow colored bacterial culture on Tripticase Soy Agar (TSA) medium

A phylogenetic tree was constructed (Fig. 2) from the 16S rRNA gene sequence together with other Arthrobacter [1, 2] homologs using MEGA 6.0 software suite [26]. The evolutionary history was inferred by using the Maximum Likelihood method based on the Tamura-Nei model [27]. The tree with the highest log likelihood (0.14495825) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. Initial tree(s) for the heuristic search were obtained automatically by applying Neighbor-Join and BioNJ algorithms to a matrix of pairwise distances estimated using the Maximum Composite Likelihood (MCL) approach, and then selecting the topology with superior log likelihood value. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. The analysis involved 13 nucleotide sequences. All positions containing gaps and missing data were eliminated. There were a total of 1553 positions in the final dataset. Evolutionary analyses were conducted in MEGA6.0 [26]. According to the 16S rRNA gene similarity, the nearest phylogenetic neighbors of Arthrobacter agilis strain L77 are Arthrobacter flavusJCM 11496T [24] (AB537168) with 97.8 %, A. tectiLMG 22282T [20] (AJ639829) with 97.13 %, A. parietisLMG 22281T [20] (AJ639830) with 97.41 %, A. subtrerraneus CH7TDSM 17585 [21] (DQ097525) with 97.66 % and A. tumbaeLMG 19501T [20] (AJ315069) with 97.68 % similarity. The 16S rRNA gene sequence also submitted to NCBI GenBank with the accession number KT804924.

Fig. 2.

Fig. 2

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Phylogenetic placements of Arthrobacter agilis strain L77 between known species of Arthrobacter genus

Extended feature descriptions

Arthrobacter agilis strain L77, a psychrotrophic bacterium, forms bright yellow color colonies (Fig. 1) on TSA medium and could grow in a pH range of 6–9 and tolerate 5 % NaCl. Growth studies showed that the isolate when incubated at 15 and 30 °C was in the exponential phase until 36 h, while at 4 °C, the exponential phase started after 24 h (Fig. 3). Freezing survival studies of Arthrobacter agilis strain L77 revealed that when the culture was initially grown at 4 °C prior to freezing at −10 and −20 °C, it showed significantly higher freezing survival rather than culture initially grown at 15 and 30 °C prior to freezing (Fig. 3).

Fig. 3.

Fig. 3

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Growth curves of Arthrobacter agilis strain L77 at three different temperatures 4, 15 and 30 °C

Exopolysaccharide production was found to be higher at lower incubation temperatures (4 or 15 °C) in comparison to the optimal growth temperature (30 °C) for Arthrobacter agilis (L77) (Fig. 4). EPS production by psychrophilic bacteria is one of the adaptations at low temperatures. The high polyhydroxyl content of EPS lowers the freezing point and ice nucleation temperature of water. In addition, EPS can trap water, nutrients and metal-ions and facilitate surface adhesion, cellular aggregation and biofilm formation and may also play a role in protecting extracellular enzymes against cold denaturation and autolysis [28, 29].

Fig. 4.

Fig. 4

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The survival of Arthrobacter agilis strain L77 subjected to freezing temperature (−10 and −20 °C) shifted from three different temperatures 4, 15 and 30 °C

Remarkable variations in terms of accumulation of various organic acids, sugars, polyols and amino acids were detected through HPLC at three different incubation temperatures (4, 15 and 30 °C) (Additional file 1: Table S1, Additional file 2: Table S2 and Fig. 5). Among the sugars, accumulation of mannitol and sorbitol was observed only at 4 °C. The amino acids expression pattern revealed that the most prominent increase was observed in the concentrations of glycine, cysteine and arginine at 4 °C (Additional file 2: Table S2). It has been reported that the cold active enzymes and efficient growth rates are used to facilitate and maintain the adequate metabolic fluxes at near freezing temperature for cold-adaptation [30]. The development of freezing tolerance by producing cryoprotectant compounds or adaptation of cytoplasmic enzymes to cold conditions for protecting cytoplasmic components is one of the strategy used by microbial cells to survive in freezing conditions as these molecules depress freezing point for the protection of cells [31].

Fig. 5.

Fig. 5

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EPS accumulation by Arthrobacter agilis strain L77 at three different temperatures 4, 15 and 30 °C

Enhanced EPS production by the psychrophilic bacteria at low temperature suggests that EPS plays an important role in desiccation protection or prevention of drying of bacterial cells from freezing temperature. It can be assumed that the strain L77 follows a cold evading strategy to thrive in freezing conditions by synthesizing various cryoprotectants (sugars, polyols and amino acids). These cryoprotectants are known to depress freezing point to evade crystallization [32].

Genome sequencing information

Genome project history

This organism was selected for sequencing on the basis of its environmental and agricultural relevance to help in plant growth and ability to provide inorganic phosphate to crops at very low temperature. It also has biogeochemical importance of producing AFCs, so helpful for soil aeration. The genome project is deposited in the online genome database (NCBI-Genome). Sequencing, assembly and annotations were performed at Division of Microbiology, Indian Agricultural Research Institute (ICAR-IARI), New Delhi, India. A summary of the project information is shown in the Table 2.

Table 2.

Genome sequencing project information for Arthrobacter agilis strain L77

MIGS ID Property Term
MIGS-31 Finishing quality Unfinished, improved high quality draft
MIGS-28 Libraries used Paired End (insert size 250 bp)
MIGS-29 Sequencing platforms Illumina MiSeq
MIGS-31.2 Fold coverage 180×
MIGS-30 Assemblers A5 pipeline v jan-2014
MIGS-32 Gene calling method Prodigal
Locus Tag RY94
Genbank ID JWSU00000000.1-10.1
Genbank Date of Release 08-Jan-2015
GOLD ID Gp0117366
BIOPROJECT PRJNA270909
MIGS 13 Source Material Identifier L77
Project relevance Bioprospecting
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Growth conditions and genomic DNA preparation

A culture of L77 was grown in Trypticase soya broth, until they reached an OD(600 nm) > 1.0. The cells were pelleted from 5 ml culture, washed thrice with TE buffer (10 mM Tris and 1 mM EDTA, pH 8.0) and the pellet was resuspended in 750 μl TE buffer. Genomic DNA was extracted from the suspended pellet using Zymo Research Fungal/Bacterial DNA MicroPrep™ following the standard protocol prescribed by the manufacturer.

Genome sequencing and assembly

The draft genome of Arthrobacter agilis strain L77 (PRJNA270909) was generated at the Division of Microbiology, ICAR-Indian Agricultural Research Institute (ICAR-IARI), New Delhi, India using Illumina [33] technology (Table 2). For this genome, we constructed and sequenced an Illumina MiSeq shotgun library which generated 1,568,654 reads totaling 321.8 Mb data. The raw fastq data was checked for quality using Fast QC [34]. Trimmomatic 0.32 [35] with Nextra adapter sequences was used to hard clip reads. Assembly of trimmed reads was carried out using a5 pipeline version 2014 [36] (Table 2). In terms of N50 and total number of scaffolds, the a5 pipeline [36] was found to be better than other genome assemblers. CONTIGuator [37] was used to improve the assembly draft. The final draft was identified as Arthrobacter agilis L77, using megablast with RDP 16S database, release 11–1 [38]. This whole-genome project (Bioproject ID: PRJNA270909) has been registered and assembled sequence data submitted at NCBI GenBank under the accession no. JWSU00000000.1-10.1. The version described in this paper is the first version.

Genome annotation

Genes were identified using Prokka 1.8 [39] based on Prodigal [40] (Table 2) as part of the Oak Ridge National Laboratory genome annotation pipeline. The predicted CDSs were further annotated on Pfam [41], and (COGs) [42]. These data sources were combined to assert a product description for each predicted protein. Non-coding genes and miscellaneous features were predicted using tRNAscan-SE [43], RNAMMer [44], Rfam [45], TMHMM [46], and signalP v4.1 [47] (Table 3).

Table 3.

Genome Statistics for Arthrobacter agilis strain L77

Attribute Value % of total
Genome size (bp) 3,608,439 100.00
DNA coding (bp) 3,224,998 89.37
DNA G + C (bp) 2,518,329 69.79
DNA scaffolds 10 100.00
Total genes 3390 100.00
Protein coding genes 3316 97.81
RNA genes 84 2.18
Pseudo genes 25 0.73
Genes in internal clusters N/A N/A
Genes with function prediction 2591 78.10
Genes assigned to COGs 2122 63.64
Genes assigned to Pfam domains 2855 85.11
Genes with signal peptides 126 5.51
Genes with transmembrane helices 852 25.6
CRISPR repeats N/A N/A
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Genome properties

The genome is 3,608,439 bp in size, which has GC content of 69.79 mol % (Table 3). There are 47 tRNA, 1 tmRNA, 6 rRNA and 20 ncRNA genes. Of the 3390 predicted genes, 3316 are protein-coding genes (CDSs). Of the total CDSs, 63.64 % represent COG functional categories and 5.51 % consist of signal peptides (Table 3). The distribution of genes into COG functional categories are presented in Table 4. The genome map (Fig. 6) was visualized by CG view server [48].

Table 4.

Number of protein coding genes of Arthrobacter agilis strain L77 associated with general COG functional categories

Code Value % agea COG category
J 184 5.54 Translation, ribosomal structure and biogenesis
A 1 0.03 RNA processing and modification
K 208 6.27 Transcription
L 109 3.28 Replication recombination and repair
B 1 0.03 Chromatin structure and dynamics
D 22 0.66 Cell cycle control, Cell division, chromosome partitioning
V 49 1.47 Defense mechanisms
T 113 3.40 Signal transduction mechanisms
M 124 3.73 Cell wall/membrane biogenesis
N 30 0.90 Cell motility
U 19 0.57 Intracellular trafficking and secretion
O 104 3.13 Posttranslational modification, protein turnover, chaperones
C 110 3.31 Energy production and conversion
G 213 6.42 Carbohydrate transport and metabolism
E 200 6.03 Amino acid transport and metabolism
F 71 2.14 Nucleotide transport and metabolism
H 114 3.43 Coenzyme transport and metabolism
I 88 2.65 Lipid transport and metabolism
P 118 3.55 Inorganic ion transport and metabolism
Q 38 1.14 Secondary metabolites biosynthesis, transport and catabolism
R 204 6.15 General function prediction only
S 166 5.00 Function unknown
– 1030 31.06 Not in COGs
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aThe total is based on the number of protein coding genes in the annotated genome

Fig. 6.

Fig. 6

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Graphical map of genome of Arthrobacter agilis strain L77. From outside to centre: RNA genes (Brown, tRNA and light purple, rRNA) and other genes are colored according to COG categories. Inner circle shows the GC skew with positive (+) as dark green and negative (−) as dark purple. GC content is indicated in black

Insights from the genome sequence

The isolate was successfully screened for lipase, amylase, protease, chitinase and β-galactosidase. Genome analysis showed two important genes pstA and pstC which are required for the translocation of phosphate across the membranes. Another important gene, PstB (an ADP binding protein), of the phosphate transport system is responsible for giving energy to the phosphate transport system of the organism. PhoR and PhoP were also found which are important for regulation of phosphate operon. PhoH like protein has a probable ATPase which is induced when phosphate level decreases. Genome annotation also predicted a putative cold shock protein which is supposed to play an important role in low temperature conditions. There are other proteins which shares evolutionary relationship with bacterial cold shock proteins such as Rhodanase and S1 RNA binding protein suggesting their role in low temperature conditions. In-depth analysis of the genome could give us better insight into mechanism of tolerance of this strain to low temperature. Other temperature responsive proteins were found such as molecular chaperone Hsp31 and glyoxalase 3 that influence the exposure of hydrophobic domains of proteins and stabilize the early unfolding under high temperature stress conditions to provide stability to the isolate in temperature stress.

Genes of heavy metal resistance were also found in the annotation. Mercuric resistance operon regulatory protein activates the mercury resistance operon in the presence of mercury thus protecting the bacteria from harmful side-effects of mercury. Mercuric reductase is also present which is responsible for conversion of Hg2+ to Hg0. copZ is a copper chaperone that replaces zinc with copper and releases copY from the DNA which is a negative regulator of copYZAB under excess copper. Gene of nitrogen regulation, nitrogen regulatory protein P-II was found that regulates the level of nitrogen by regulating glutamine. When the ratio of glutamine to 2-ketoglutarate decreases, uridine is added on a tyrosine of P-II to form P-II-UMP which in turn deadenylates glutamine synthase resulting in its activation. Putative genes coding for these activities were identified in the genome based on annotation (Table 5).

Table 5.

Candidate genes coding for putative lipase, amylase, chitinase, protease, β-galactosidase, phosphate transport regulation, cold shock proteins, chaperons and heavy metal resistance activities identified in Arthrobacter agilis strain L77 draft genome

Putative Gene Annotation Size (aa)
Lipase
 ABAGL_00531 GDSL-like Lipase/Acylhydrolase 262
 ABAGL_00732 Lipase 1 precursor 288
 ABAGL_00875 GDSL-like Lipase/Acylhydrolase 267
 ABAGL_01161 Lipase 1 precursor 350
 ABAGL_03217 GDSL-like Lipase/Acylhydrolase 272
Amylase
 ABAGL_00299 Glucose-resistance amylase regulator 338
 ABAGL_01452 Glucose-resistance amylase regulator 336
 ABAGL_01652 Trehalose synthase/amylase TreS 588
 ABAGL_01737 Alpha-amylase precursor 905
 ABAGL_01923 Alpha-amylase/pullulanase 257
 ABAGL_01950 Glucose-resistance amylase regulator 327
Chitinase
 ABAGL_01394 putative bifunctional chitinase/lysozyme precursor 520
 ABAGL_01777 Chitinase 400
Protease
 ABAGL_00100 Putative cysteine protease YraA 188
 ABAGL_00190 Flp pilus assembly protein, protease CpaA 207
 ABAGL_00447 Lon protease 364
 ABAGL_00456 Putative serine protease HtrA 496
 ABAGL_00667 Serine proteasec 401
 ABAGL_00940 CAAX amino terminal protease self- immunity 268
 ABAGL_00971 CAAX amino terminal protease self- immunity 247
 ABAGL_01091 Serine protease Do-like HtrA 366
 ABAGL_01213 Rhomboid protease GluP 291
 ABAGL_01289 ATP-dependent zinc metalloprotease FtsH 689
 ABAGL_01302 Putative ATP-dependent Clp protease ATP-binding subunit 835
 ABAGL_01392 CAAX amino terminal protease self- immunity 266
 ABAGL_01505 Minor extracellular protease vpr precursor 1059
 ABAGL_01669 Flp pilus assembly protein, protease CpaA 168
 ABAGL_01755 CAAX amino terminal protease self- immunity 326
 ABAGL_02020 Putative serine protease HtrA 310
 ABAGL_02206 Putative metalloprotease 303
 ABAGL_02449 Putative zinc metalloproteasec/MT2700 388
 ABAGL_02467 Modulator of FtsH protease HflK 310
 ABAGL_02638 ATP-dependent Clp protease ATP-binding subunit ClpX 430
 ABAGL_02639 ATP-dependent Clp protease proteolytic subunit 1 224
 ABAGL_02640 ATP-dependent Clp protease proteolytic subunit 2 208
 ABAGL_02862 ATP-dependent Clp protease adaptor protein ClpS 105
 ABAGL_02923 ATP-dependent zinc metalloprotease FtsH 438
 ABAGL_03163 Serine protease inhibitor-like protein 389
 ABAGL_03211 CAAX amino terminal protease self- immunity 267
 ABAGL_03271 Metalloprotease MmpA 447
 ABAGL_00551 Protease PrtS precursor 355
 ABAGL_00739 Protease 2 734
 ABAGL_01958 Protease synthase and sporulation negative regulatory protein 215
 ABAGL_02571 Protease PrsW 425
 ABAGL_03295 Protease 3 precursor 455
β-galactosidase
 ABAGL_00260 β-galactosidase bgaB 667
 ABAGL_00292 β-galactosidase 687
 ABAGL_01083 β-galactosidase precursor 708
Phosphate Transport Regulation
 ABAGL_01317 Phosphate transport system permease protein PstA 310
 ABAGL_01318 Phosphate import ATP-binding protein PstB 367
 ABAGL_01316 Phosphate transport system permease protein PstC 259
 ABAGL_00191 Alkaline phosphatase synthesis sensor protein PhoR 544
 ABAGL_03137 Alkaline phosphatase synthesis sensor protein PhoR 555
 ABAGL_01671 PhoH-like protein 443
 ABAGL_02530 PhoH-like protein 344
Cold shock Proteins
 ABAGL_01978 putative cold shock protein A 67
Chaperons
 ABAGL_01554 Molecular chaperone Hsp31 and glyoxalase 3 255
 ABAGL_01067 Copper chaperone CopZ 74
Heavy Metal Resistance
 ABAGL_02628 Mercuric resistance operon regulatory protein 134
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Conclusions

The 3.6 Mb draft genome of Arthrobacter agilis strain L77 was assembled and annotated. The isolate was successfully screened for production of EPS and AFCs with potential application in biotechnology. The candidate genes coding for hydrolytic enzymes and cold shock proteins were identified in the genome. Arthrobacter agilis strain L77 will serve as a source for antifreeze proteins, functional enzymes and other bioactive molecules in future bioprospecting projects.

Acknowledgments

The authors are grateful to the National Agricultural Innovation Project (NAIP), Indian Council of Agricultural Research, Govt. of India, New Delhi and Division of Microbiology, ICAR-Indian Agricultural Research Institute (IARI), Pusa, New Delhi and for providing the financial support facilities, to undertake the investigations.

Author’s contributions

RNS and SGa equally contributed to the work. RNS carried out the sample collection, participated in the strain identification, sequence alignment, assembly and annotation analysis and drafted the manuscript. SGa participated in the sequence assembly and annotation analysis. ANY and SGu carried out the bacterial isolation and performed the physiological assays. PG did the initial sequence assembly of the raw data. RK participated in sample collection and sequencing of 16S rRNA gene. AKS conceived of the study, and participated in its design, coordination and helped to finalize the manuscript. All authors read and approved the final manuscript.

Competing interests

The authors declare that they have no competing interests.

Abbreviations

AFCs

Anti-freeze compounds

EPS

Exopolysaccharides

Additional files

Additional file 1: Table S1. (13.5KB, docx)

Quantitative analysis of organic acid and sugars/polyols from Arthrobacter agilis strain L77 by HPLC. (DOCX 13 kb)

Additional file 2: Table S2. (15.4KB, docx)

Quantitative analysis of amino acids content of Arthrobacter agilis strain L77 by HPLC. (DOCX 15 kb)

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