Heterologous expression of an <i>in planta</i>-upregulated gene cluster in the wheat pathogen <i>Parastagonospora nodorum</i> establishes the biosynthesis of methylene-bridged depsides.
Nguyen, Hera T.; Sbaraini, Nicolau; Bracegirdle, Joe; Smithers, Luke; Moggach, Stephen A.; Vuong, Daniel; Lacey, Ernest; Solomon, Peter S.; Haywood, Joel; Piggott, Andrew M.; Chooi, Yit-Heng; Chemical science 16 (2025) 21852–21861; doi:10.1039/d5sc05442g

data_1576016
_publ_section_title
;
 Heterologous expression of an <i>in planta</i>-upregulated gene cluster
 in the wheat pathogen <i>Parastagonospora nodorum</i> establishes the
 biosynthesis of methylene-bridged depsides.
;
_journal_issue                   46
_journal_name_full               'Chemical science'
_journal_page_first              21852
_journal_page_last               21861
_journal_paper_doi               10.1039/d5sc05442g
_journal_volume                  16
_journal_year                    2025
_chemical_formula_moiety         'C20 H22 O8'
_chemical_formula_sum            'C20 H22 O8'
_chemical_formula_weight         390.37
_space_group_crystal_system      triclinic
_space_group_IT_number           2
_space_group_name_Hall           '-P 1'
_space_group_name_H-M_alt        'P -1'
_atom_sites_solution_hydrogens   geom
_atom_sites_solution_primary     dual
_audit_creation_date             2025-05-19
_audit_creation_method
;
Olex2 1.5
(compiled 2025.04.05 svn.rf049d14d for OlexSys, GUI svn.r7224)
;
_audit_update_record
;
2025-07-16 deposited with the CCDC.	2025-10-14 downloaded from the CCDC.
;
_cell_angle_alpha                90.071(2)
_cell_angle_beta                 99.076(3)
_cell_angle_gamma                111.840(4)
_cell_formula_units_Z            2
_cell_length_a                   7.3057(3)
_cell_length_b                   8.9953(4)
_cell_length_c                   14.6402(3)
_cell_measurement_reflns_used    8197
_cell_measurement_temperature    100.02(10)
_cell_measurement_theta_max      73.3890
_cell_measurement_theta_min      3.0430
_cell_volume                     879.99(6)
_computing_cell_refinement       'CrysAlisPro 1.171.43.143a (Rigaku OD, 2024)'
_computing_data_collection
'CrysAlisPro system (CCD 43.143a 64-bit (release 25-10-2024))'
_computing_data_reduction        'CrysAlisPro 1.171.43.143a (Rigaku OD, 2024)'
_computing_molecular_graphics    'Olex2 1.5 (Dolomanov et al., 2009)'
_computing_publication_material  'Olex2 1.5 (Dolomanov et al., 2009)'
_computing_structure_refinement  'XL  (Sheldrick, 2008)'
_computing_structure_solution    'SHELXT 2018/2 (Sheldrick, 2018)'
_diffrn_ambient_environment      N~2~
_diffrn_ambient_temperature      100.02(10)
_diffrn_detector                 'Hybrid Pixel Array Detector'
_diffrn_detector_area_resol_mean 10.0000
_diffrn_detector_type            HyPix
_diffrn_measured_fraction_theta_full 0.997
_diffrn_measured_fraction_theta_max 0.955
_diffrn_measurement_details
;
List of Runs (angles in degrees, time in seconds):

  # Type    Start    End  Width  t~exp~   \w     \q     \k     \f   Frames
  1  \w    -94.00 -14.00   0.50    2.50    --  -46.32 127.00-109.00  160
  2  \w   -104.00 -23.00   0.50    2.50    --  -46.32 144.00  59.00  162
  3  \w   -118.00 -60.00   0.50    2.50    --  -46.32-144.00   5.00  116
  4  \w     61.00 119.00   0.50    2.50    --   46.32 144.00  59.00  116
  5  \w     23.00 104.00   0.50    2.50    --   46.32-144.00   5.00  162
  6  \w     14.00  94.00   0.50    2.50    --   46.32-127.00 173.00  160
  7  \w   -125.00 -30.00   0.50    9.00    -- -102.29 144.00  59.00  190
  8  \w   -131.00 -95.00   0.50    9.00    -- -102.29-127.00 173.00   72
  9  \w   -140.00-114.00   0.50    9.00    -- -102.29-144.00   5.00   52
 10  \w    -99.00 -58.00   0.50    9.00    -- -102.29 127.00-109.00   82
 11  \w     76.00 109.00   0.50    9.00    --  102.29-144.00   5.00   66
 12  \w     98.00 123.00   0.50    9.00    --  102.29 127.00-109.00   50
 13  \w     79.00 116.00   0.50    9.00    --  102.29 144.00  59.00   74
 14  \w     35.00 112.00   0.50    9.00    --  102.29-127.00 173.00  154
 15  \w    -57.00 -32.00   0.50    2.50    --   14.00 164.00-101.55   50
 16  \w    -17.00   8.00   0.50    2.50    --   14.00  97.00  27.50   50
 17  \w      3.00  30.00   0.50    2.50    --   30.00  94.00  63.15   54
 18  \w     13.00  38.00   0.50    2.50    --   42.00  90.00-134.03   50
 19  \w     12.00  37.00   0.50    2.50    --   42.00  92.00  80.03   50
 20  \w    -86.00 -40.00   0.50    2.50    --  -46.32 -19.00 120.00   92
 21  \w    -56.00 -30.00   0.50    2.50    --  -46.32 -77.00-180.00   52
 22  \w    -74.00 -42.00   0.50    2.50    --  -46.32 -38.00-150.00   64
 23  \w     35.00  74.00   0.50    2.50    --   46.32  38.00 -90.00   78
 24  \w     28.00  66.00   0.50    2.50    --   50.00  76.00  75.78   76
 25  \w     27.00  56.00   0.50    2.50    --   50.00  77.00 -24.02   58
 26  \w     32.00  65.00   0.50    2.50    --   54.00  76.00  -8.88   66
 27  \w     32.00  65.00   0.50    2.50    --   54.00  76.00 112.51   66
 28  \w     33.00  59.00   0.50    2.50    --   54.00  75.00-166.30   52
 29  \w     80.00 106.00   0.50    2.50    --   54.00  74.00 -57.89   52
 30  \w     36.00  62.00   0.50    2.50    --   54.00  74.00 -57.89   52
 31  \w     32.00  58.00   0.50    2.50    --   54.00  77.00  19.54   52
 32  \w     37.00  63.00   0.50    2.50    --   58.00  74.00 -92.64   52
 33  \w     40.00  66.00   0.50    2.50    --   62.00  75.00 134.15   52
 34  \w     42.00  68.00   0.50    2.50    --   62.00  73.00 154.54   52
 35  \w    -74.00 -48.00   0.50    2.50    --  -66.00 -68.00-161.43   52
 36  \w    -71.00 -46.00   0.50    2.50    --  -66.00 -69.00 128.60   50
 37  \w    -77.00 -51.00   0.50    2.50    --  -66.00 -62.00 -81.88   52
 38  \w     45.00  70.00   0.50    2.50    --   66.00  73.00  41.83   50
 39  \w     45.00  71.00   0.50    2.50    --   66.00  76.00 110.03   52
 40  \w    -77.00 -51.00   0.50    2.50    --  -70.00 -67.00 -33.77   52
 41  \w    -78.00 -52.00   0.50    2.50    --  -74.00 -75.00 -80.09   52
 42  \w    -84.00 -58.00   0.50    2.50    --  -74.00 -61.00 -52.71   52
 43  \w    -91.00 -57.00   0.50    2.50    --  -74.00 -61.00-138.92   68
 44  \w    -82.00 -54.00   0.50    2.50    --  -74.00 -74.00 -89.53   56
 45  \w    -84.00 -52.00   0.50    2.50    --  -74.00 -74.00-167.40   64
 46  \w    -87.00 -59.00   0.50    2.50    --  -74.00 -59.00  -7.04   56
 47  \w    -86.00 -60.00   0.50    2.50    --  -74.00 -59.00  45.71   52
 48  \w    -77.00 -52.00   0.50    2.50    --  -74.00 -74.00-103.60   50
 49  \w    -82.00 -57.00   0.50    9.00    --  -78.00 -73.00-104.77   50
 50  \w   -128.00-102.00   0.50    9.00    --  -78.00 -73.00-104.77   52
 51  \w    -84.00 -58.00   0.50    9.00    --  -78.00 -73.00  63.23   52
 52  \w    -93.00 -61.00   0.50    9.00    --  -78.00 -71.00  39.17   64
 53  \w    -90.00 -65.00   0.50    9.00    --  -82.00 -61.00 -29.74   50
 54  \w    -98.00 -64.00   0.50    9.00    --  -82.00 -64.00 156.54   68
 55  \w   -100.00 -60.00   0.50    9.00    --  -82.00 -74.00 131.90   80
 56  \w    -88.00 -59.00   0.50    9.00    --  -82.00 -76.00 120.78   58
 57  \w   -118.00 -92.00   0.50    9.00    --  -86.00 -73.00-108.34   52
 58  \w   -148.00-122.00   0.50    9.00    --  -86.00 -76.00-123.60   52
 59  \w   -116.00 -63.00   0.50    9.00    --  -86.00 -76.00-123.60  106
 60  \w    -91.00 -66.00   0.50    9.00    --  -86.00 -71.00  -2.00   50
 61  \w   -103.00 -71.00   0.50    9.00    --  -86.00 -63.00  89.83   64
 62  \w    -91.00 -65.00   0.50    9.00    --  -86.00 -72.00  38.70   52
 63  \w    -89.00 -64.00   0.50    9.00    --  -86.00 -74.00 145.86   50
 64  \w    -90.00 -65.00   0.50    9.00    --  -86.00 -73.00-108.34   50
 65  \w   -162.00-101.00   0.50    9.00    --  -90.00 -63.00 129.94  122
 66  \w    -94.00 -69.00   0.50    9.00    --  -90.00 -73.00-153.64   50
 67  \w   -139.00-113.00   0.50    9.00    --  -90.00 -73.00-153.64   52
 68  \w    -94.00 -69.00   0.50    9.00    --  -90.00 -73.00 -36.03   50
 69  \w   -103.00 -74.00   0.50    9.00    --  -90.00 -65.00 -99.38   58
 70  \w   -162.00-105.00   0.50    9.00    --  -90.00 -73.00 -36.03  114
 71  \w   -105.00 -75.00   0.50    9.00    --  -90.00 -55.00 -93.15   60
 72  \w   -121.00 -76.00   0.50    9.00    --  -90.00 -57.00 -39.33   90
 73  \w   -134.00 -76.00   0.50    9.00    --  -94.00 -66.00  86.25  116
 74  \w   -101.00 -76.00   0.50    9.00    --  -94.00 -65.00  91.41   50
 75  \w   -166.00-121.00   0.50    9.00    --  -94.00 -65.00  91.41   90
 76  \w   -164.00 -73.00   0.50    9.00    --  -94.00 -76.00 166.02  182
 77  \w   -119.00 -78.00   0.50    9.00    --  -94.00 -64.00-147.24   82
 78  \w   -110.00 -77.00   0.50    9.00    --  -94.00 -64.00  24.41   66
 79  \w   -160.00-127.00   0.50    9.00    --  -94.00 -64.00  24.41   66
 80  \w   -146.00 -79.00   0.50    9.00    --  -98.00 -68.00-159.53  134
 81  \w   -133.00 -80.00   0.50    9.00    --  -98.00 -64.00  63.87  106
 82  \w   -167.00-141.00   0.50    9.00    --  -98.00 -64.00  63.87   52
 83  \w   -126.00 -80.00   0.50    9.00    --  -98.00 -64.00 -61.98   92
 84  \w   -142.00 -84.00   0.50    9.00    -- -102.29 -66.00  -3.85  116
 85  \w   -115.00 -80.00   0.50    9.00    -- -102.29 -78.00  22.47   70
 86  \w    -63.00 -30.00   0.50    9.00    -- -102.29  61.00 -90.00   66
 87  \w   -168.00-121.00   0.50    9.00    -- -102.29 -78.00  22.47   94
 88  \w    -63.00 -32.00   0.50    9.00    -- -102.29  61.00-180.00   62
 89  \w   -175.00 -79.00   0.50    9.00    -- -102.29 -79.00-100.49  192
 90  \w   -104.00 -79.00   0.50    9.00    -- -102.29 -78.00  58.76   50
 91  \w   -169.00-115.00   0.50    9.00    -- -102.29 -78.00  58.76  108
 92  \w   -108.00 -82.00   0.50    9.00    -- -102.29 -75.00 -57.69   52
 93  \w   -170.00-118.00   0.50    9.00    -- -102.29 -75.00 -57.69  104
 94  \w    -73.00 -33.00   0.50    9.00    -- -102.29  61.00-150.00   80
 95  \w   -148.00 -80.00   0.50    9.00    -- -102.29 -75.00-163.97  136
 96  \w   -105.00 -30.00   0.50    9.00    -- -102.29  77.00 -90.00  150
 97  \w     73.00 104.00   0.50    9.00    --  102.29 -94.00  60.00   62
 98  \w     57.00 105.00   0.50    9.00    --  102.29 -94.00   0.00   96
 99  \w     30.00  67.00   0.50    9.00    --  102.29 -94.00  60.00   74
100  \w     78.00 103.00   0.50    9.00    --  102.29 -77.00 -30.00   50
101  \w     31.00  62.00   0.50    9.00    --  102.29 -77.00 -30.00   62
102  \w     32.00  68.00   0.50    9.00    --  102.29 -61.00  60.00   72
103  \w     31.00  69.00   0.50    9.00    --  102.29 -61.00 -30.00   76
104  \w     70.00 104.00   0.50    9.00    --  102.29 -94.00-180.00   68
;
_diffrn_measurement_device       'four-circle diffractometer'
_diffrn_measurement_device_type
'XtaLAB Synergy, Single source at home/near, HyPix'
_diffrn_measurement_method       '\w scans'
_diffrn_orient_matrix_type
'CrysAlisPro convention (1999,Acta A55,543-557)'
_diffrn_orient_matrix_UB_11      -0.1279608000
_diffrn_orient_matrix_UB_12      0.0165208000
_diffrn_orient_matrix_UB_13      -0.0907540000
_diffrn_orient_matrix_UB_21      0.1483006000
_diffrn_orient_matrix_UB_22      -0.0267944000
_diffrn_orient_matrix_UB_23      -0.0558348000
_diffrn_orient_matrix_UB_31      -0.1215995000
_diffrn_orient_matrix_UB_32      -0.1822027000
_diffrn_orient_matrix_UB_33      -0.0071023000
_diffrn_radiation_monochromator  mirror
_diffrn_radiation_probe          x-ray
_diffrn_radiation_type           'Cu K\a'
_diffrn_radiation_wavelength     1.54184
_diffrn_reflns_av_R_equivalents  0.0661
_diffrn_reflns_av_unetI/netI     0.0344
_diffrn_reflns_Laue_measured_fraction_full 0.997
_diffrn_reflns_Laue_measured_fraction_max 0.955
_diffrn_reflns_limit_h_max       9
_diffrn_reflns_limit_h_min       -9
_diffrn_reflns_limit_k_max       10
_diffrn_reflns_limit_k_min       -11
_diffrn_reflns_limit_l_max       18
_diffrn_reflns_limit_l_min       -18
_diffrn_reflns_number            25212
_diffrn_reflns_point_group_measured_fraction_full 0.997
_diffrn_reflns_point_group_measured_fraction_max 0.955
_diffrn_reflns_theta_full        67.684
_diffrn_reflns_theta_max         75.286
_diffrn_reflns_theta_min         3.063
_diffrn_source                   'micro-focus sealed X-ray tube'
_diffrn_source_type              'PhotonJet (Cu) X-ray Source'
_exptl_absorpt_coefficient_mu    0.965
_exptl_absorpt_correction_T_max  1.00000
_exptl_absorpt_correction_T_min  0.72906
_exptl_absorpt_correction_type   multi-scan
_exptl_absorpt_process_details
;
CrysAlisPro 1.171.43.143a (Rigaku Oxford Diffraction, 2024)
Empirical absorption correction using spherical harmonics,
implemented in SCALE3 ABSPACK scaling algorithm.
;
_exptl_crystal_colour            'clear light colourless'
_exptl_crystal_density_diffrn    1.473
_exptl_crystal_description       block
_exptl_crystal_F_000             412
_exptl_crystal_size_max          0.08
_exptl_crystal_size_mid          0.07
_exptl_crystal_size_min          0.05
_refine_diff_density_max         0.421
_refine_diff_density_min         -0.334
_refine_diff_density_rms         0.072
_refine_ls_extinction_method     none
_refine_ls_goodness_of_fit_ref   1.072
_refine_ls_hydrogen_treatment    constr
_refine_ls_matrix_type           full
_refine_ls_number_parameters     262
_refine_ls_number_reflns         3473
_refine_ls_number_restraints     0
_refine_ls_restrained_S_all      1.072
_refine_ls_R_factor_all          0.0743
_refine_ls_R_factor_gt           0.0652
_refine_ls_shift/su_max          0.000
_refine_ls_shift/su_mean         0.000
_refine_ls_structure_factor_coef Fsqd
_refine_ls_weighting_details
'w=1/[\s^2^(Fo^2^)+(0.1119P)^2^+0.5726P] where P=(Fo^2^+2Fc^2^)/3'
_refine_ls_weighting_scheme      calc
_refine_ls_wR_factor_gt          0.1789
_refine_ls_wR_factor_ref         0.1873
_reflns_Friedel_coverage         0.000
_reflns_number_gt                3035
_reflns_number_total             3473
_reflns_threshold_expression     'I > 2\s(I)'
_cod_data_source_file            d5sc05442g2.cif
_cod_data_source_block           hcjf8
_cod_depositor_comments
'Adding full bibliography for 1576015--1576018.cif.'
_cod_database_code               1576016
_shelx_shelxl_version_number     2019/1
_chemical_oxdiff_formula         'C19 H12 N2'
_chemical_oxdiff_usercomment     'Hera for Heng'
_cell_oxdiff_angle_alpha         90.071(2)
_cell_oxdiff_angle_beta          99.076(3)
_cell_oxdiff_angle_gamma         111.840(4)
_cell_oxdiff_length_a            7.3057(3)
_cell_oxdiff_length_b            8.9953(4)
_cell_oxdiff_length_c            14.6402(3)
_cell_oxdiff_measurement_reflns_used 8197
_cell_oxdiff_volume              879.98(5)
_shelx_estimated_absorpt_t_max   0.953
_shelx_estimated_absorpt_t_min   0.927
_olex2_refinement_description
;
1. Fixed Uiso
 At 1.2 times of:
  All C(H) groups, All C(H,H) groups
 At 1.5 times of:
  All C(H,H,H) groups, All O(H) groups
2.a Secondary CH2 refined with riding coordinates:
 C00M(H00d,H00e)
2.b Aromatic/amide H refined with riding coordinates:
 C00H(H00H)
2.c Idealised Me refined as rotating group:
 C00L(H00a,H00b,H00c), C00P(H00f,H00g,H00i), C00Q(H00j,H00k,H00l), C00R(H00m,
 H00n,H00o), C00S(H00p,H00q,H00r)
2.d Idealised tetrahedral OH refined as rotating group:
 O002(H002), O004(H004), O005(H005), O008(H008)
;
_shelx_res_checksum              59569
_olex2_submission_special_instructions 'No special instructions were received'
_oxdiff_exptl_absorpt_empirical_details
;
Empirical correction (ABSPACK) includes:
- Absorption correction using spherical harmonics
- Frame scaling
;
_oxdiff_exptl_absorpt_empirical_full_max 1.651
_oxdiff_exptl_absorpt_empirical_full_min 0.655